Supplementary Fig. 1 a Donor # 28-IL2RB-z 28- IL2RB-z Control MFI: 1866 1394 612 1297 11 MFI: 1794 1294 566 1225 11 MFI: 1433 1157 327 1117 98.5 MFI: 1463 1224 352 111 126 MFI: 1441 1232 497 1268 151 MFI: 1344 1147 679 174 111 1 2 3 4 Protein L / streptavidin-pe 5 6 NGFR (Pacific Blue) P=.9 * 24 P=.8 18 * 12 6 Control 28-ΔIL2RBz 28-IL2RBz Mean fluorescence inteity of CAR expression b Supplementary Fig. 1. Surface expression of CAR cotructs. (a, b) Peripheral blood CD3+ T cells were individually traduced with a CAR gene linked to the truncated nerve growth factor receptor ( NGFR) or control NGFR and stained with biotin-labeled protein L and streptavidin-pe to analyze the surface expression of the CARs. FACS plots obtained from six different donor-derived T cells (a) and the mean fluorescence inteity (MFI) in the CD8+ NGFR+ T cell population (b) are shown (n=6 different donor samples). Horizontal lines indicate mean values. The MFI among different CAR cotructs was analyzed with repeated measures one-way ANOVA with Tukey s multiple compariso test; F=11.5; degree of freedom=23). * P<.1, not significant.
Supplementary Fig. 2 a 28-IL2RB-z z Control MFI: 317 2839 763 2481 244 CD69 (PE) b MFI: 122 482 perk (Alexa Fluor 647) MFI: 117 441 MFI: 995 476 MFI: 1111 439 28-IL2RBz MFI: 764 427 MFI: 741 41 MFI: 178 461 MFI: 118 44 c Mean fluorescence inteity of perk Mean fluorescence inteity of pakt P=.5 P=.45 n.s. pakt (Alexa Fluor 647) NALM-6 K562 28-IL2RBz 28- IL2RBz 28-IL2RBz 28- IL2RBz NALM-6 K562 Supplementary Fig. 2. Comparison of CAR signaling upon antigen recognition. (a) Peripheral blood CD3 + T cells were individually traduced with a CAR gene linked to the truncated nerve growth factor receptor ( NGFR) or control NGFR. CD69 expression in the CAR-traduced T cells was analyzed 24 hours after stimulation with the CD19 + cell line NALM-6. Representative FACS plots of three experiments are shown. (b, c) T cells traduced with the indicated CAR genes were stimulated with NALM-6 (CD19 + ) or K562 (CD19 - ) for 2 hours after resting overnight in cytokine-free media. Phosphorylated ERK and Akt within the CD8 + NGFR + T cell population were analyzed with intracellular flow cytometry. Representative FACS plots (d) and the MFI (e) within the CD8 + NGFR + T cell population are shown (n=4 different donor samples; repeated measures one-way ANOVA with Tukey s multiple compariso test; F=35.54 for perk and F=25.86 for pakt; degree of freedom=15 for each comparison). Horizontal lines represent mean values., not significant.
Supplementary Fig. 3 a NALM-6 K562 28-IL2RB-z z 28-IL2RB-z z IL-2 IL-21 pstat3 STAT3 pstat5 STAT5 -actin b 6 4 2 pstat3 / STAT3 pstat5 / STAT5 Supplementary Fig. 3. JAK-STAT signaling induced by CAR-T cells. (a, b) T cells traduced with the indicated CAR genes shown in Fig. 1a were stimulated with NALM-6 or K562 after resting in cytokine-free media overnight. The NGFR + T cells were isolated 2 hours after stimulation, and phosphorylated or total STAT3/STAT5 was detected by immunoblotting. T cells mock-traduced and treated with IL-2 (3 IU/ml) or IL-21 (5 ng/ml) for 3 minutes were used as a control. Representative blots of three independent experiments (a) and the quantified band inteity (b) are shown. The ratio of phosphorylated to total protei was calculated and normalized to that of the CAR-T cells stimulated with NALM-6 (n=3). Horizontal lines indicate mean values ± s.d.
Supplementary Fig. 4 a 28-IL2RBz 28-IL2RBz NGFR (FITC) CD8 (Pacific Blue) 18.6 33.9 22.3 3.1 19.3 28.7 19.5 32.5 2.4 26.3 19.5 28.8 17.9 25.7 18.2 29.1 NGFR + 437 415 438 1291 862 888 48 171 NGFR - 384 583 384 51 128 874 391 166 b Mean fluorescence inteity pstat3 (Alexa Fluor 647) 15 1 5 pstat3 P=.3 NGFR + - + - + - + - Mean fluorescence inteity NGFR pstat5 (Alexa Fluor 647) pstat5 P<.1 + - + - + - + - 28-IL2RBz 28-IL2RBz 24 16 8 c d 3 pstat3/stat3 2 pstat3 STAT3 NGFR + NGFR - Relative band inteity 1 pstat5/stat5 pstat5 STAT5 -actin Supplementary Fig. 4. JAK-STAT pathway activation in CAR + and CAR - T cells. (a-d) CAR-T cells were stimulated with NALM-6 after resting overnight in cytokine-free media. The phosphorylation of STAT3 and STAT5 was analyzed 2 hours after stimulation by flow cytometry (a and b) and immunoblotting (c and d). (a) Representative FACS plots from the samples presented in Fig. 1b. (b) Mean fluorescence inteity of individual samples (n=4; paired two-tailed t-test; t=.2 for -pstat3, t=1.29 for -pstat3, t=8.97 for z -pstat3, t=.74 for -pstat5, t=4.61 for -pstat5, t=17.99 for 28- IL2RB-z -pstat5, degree of freedom=3). Horizontal lines indicate mean values. For immunoblotting, NGFR +/- CD8 + T cells were isolated after stimulation by flow cytometry. Representative blots of four experiments (c) and the quantified band inteity normalized to that in CAR-T cells (d) are shown. Horizontal lines indicate mean values ± s.d., not significant. 28-IL2RBz 28- IL2RBz 28-IL2RBz NGFR + NGFR - 28- IL2RBz
Supplementary Fig. 5 a 28- IL2RB (FLSL)-z z b 28- IL2RB (FLSL) -z z pstat3 MFI pstat3 (Alexa Fluor 647) 1257 118 597 STAT3 pstat5 STAT5 -actin MFI pstat5 (Alexa Fluor 647) 191 1198 1837 c Relative ratio of phosphorylated/total protein 28- IL2RBz (Y XXQ) 28- IL2RB (FLSL)- z (Y XXQ) 28- IL2RB-z 28- IL2RBz (Y XXQ) 28- IL2RB (FLSL)- z (Y XXQ) 28- IL2RB-z pstat3/stat3 pstat5/stat5 Supplementary Fig. 5. Differential roles of tyrosine residues in inducing STAT3 and STAT5 phosphorylation. (a-c) T cells traduced with the indicated CARs were stimulated with NALM-6. The phosphorylation levels of STAT3 and STAT5 were analyzed 2 hours after stimulation by intracellular flow cytometry (a) or immunoblotting (b and c). Immunoblotting was performed three times, and the quantified band inteity normalized to that of the z CAR-T cells is shown (c). Horizontal lines indicate mean values ± s.d.
Supplementary Fig. 6 a Fold expaion * CD4 + P=.4 * P=.1 * Fold expaion * * CD8 + * P=.1 P=.3 P=.1 * * * * 28- IL2RBz 28- IL2RB (FLSL)-z 28- IL2RB-z 28- IL2RBz 28- IL2RB (FLSL)-z 28- IL2RB-z b 28- IL2RB (FLSL)-z z CD4 + MFI 3426 3291 2615 32 376 CD8 + CFSE MFI 288 2658 2137 271 2769 Supplementary Fig. 6. Proliferation of CAR-T cells. (a) T cells traduced with the indicated CAR genes were stimulated with NALM-6 or K562 and cultured in the absence of exogenous cytokines. Fold expaion of the CAR-T cells within CD4 + or CD8 + T cell population was analyzed 7 days after stimulation (n=8 different donor samples; repeated measures oneway ANOVA with Tukey s multiple compariso test for the NALM-6 data, F=4.47 for CD4 + T cells, F=3.84 for CD8 + T cells, degree of freedom=39; two-tailed paired t-test for comparison between the NALM-6 and K562 data in the individual CAR-T cells, t=6.79 (CD4 + -), t=5.32 (CD4+-), t=5.97 (CD4 + -z ), t=5.58 (CD4 + -28- IL2RB (FLSL)-z ), t=8.29 (CD4 + -z), t=8.73 (CD8 + -), t=7.36 (CD8 + -), t=1.8 (CD8 + -z ), t=7.45 (CD8 + -28- IL2RB (FLSL)-z ), t=7.93 (CD8 + -z), degree of freedom=7). Horizontal lines denote mean values. * P<.1. (b) CAR-T cells were labeled with carboxyfluorescein succinimidyl ester (CFSE) and stimulated with NALM-6. CFSE dilution within CD4 + /CD8 + NGFR + T cell population was analyzed three days after the stimulation. Representative FACS plots of four samples are shown.
Supplementary Fig. 7 CD62L (PE) 29.6 28- IL2RBz 16.9 3.3 3.4 18.9 35.8 23.8 22. 28- IL2RB (FLSL)-z 31. 25.1 32.7 28- IL2RB-z 34.3 18.4 19.1 24.4 CD45RA (FITC) 25.5 18.5 31.1 33.3 39.4 25.5 29.3 23.6 28. 28.1 8.8 13.5 9.7 9.9 8. CD27 (APC-Cy7) CCR7 (Pacific Blue) 58.7 16.3 62.9 26. 5.4 26.1 55.5 24.2 51. 19.9 24.5.49 1.7.32 23..46 19.8.51 27.8 1.29 49.4 24. 49.4 31.2 56.5 29.4 46.6 39.4 48.2 27.9 18.5 8.7 12.1 7.25 9.75 4.39 7.63 6.44 16.3 7.64 6.2 12.5 58.6 21.7 61.9 23.7 64. 21.7 58.8 16.7 23. 4.26 15.1 4.59 11.5 2.81 11.3 3. 2. 4.54 CD28 (APC) CCR7 (Pacific Blue) CD28 (APC) CD45RA (FITC) CD62L (PE) CCR7 (Pacific Blue) 6.4 66.2 69.7 76.4 66.7 39.4 33.3 29.7 22.9 33.2 CD95 (PerCP/Cy5.5) Supplementary Fig. 7. Expression of memory T cell markers in the CAR-traduced T cells. The expression of CD45RA, CD62L, CCR7, CD27, CD28 and CD95 was analyzed in the CAR-traduced CD8+ T cells 7 days after stimulation with NALM-6. Representative FACS plots of the nine different donor samples presented in Fig. 2d are shown.
Supplementary Fig. 8 a c MFI: 1235 471 129 pstat3 (Alexa Fluor 647) CD45RA + CD62L + CCR7 + cells within CD8 + CAR-T cells (%) 1 8 6 4 2 DMSO P=.3 S3I-21 MFI: 251 28 798 pstat5 (Alexa Fluor 647) Pimozide S3I-21+ Pimozide DMSO S3I-21 Pimozide b Mean fluorescence inteity 25 2 15 1 5 P=.12 DMSO S3I-21 pstat3 Pimozide DMSO P=.4 S3I-21 pstat5 Pimozide Supplementary Fig. 8. Specific inhibition of STAT3 and STAT5 phosphorylation. (a, b) The z CAR-T cells were stimulated with NALM-6 in the presence or absence of 25 M of S3I-21 or 5 M of pimozide and were analyzed for the phosphorylation of STAT3 and STAT5 in the CD8 + NGFR + T cell population 12 minutes after the stimulation. Representative FACS plots (a) and mean fluorescence inteity of pstat3 and pstat5 are shown (b, n=4 different donor samples; repeated measures one-way ANOVA with Tukey s multiple compariso test; F=54.54 for pstat3 and F=115.8 for pstat5; degree of freedom=11). (c) The z CAR-traduced T cells were stimulated with NALM-6 and were treated with 25 M of S3I-21 and/or 5 M of pimozide for three days. The frequency of CD45RA + CD62L + CCR7 + cells within the CD8 + NGFR + T cell population was analyzed on day 7 (n=7 different donor samples, repeated measures one-way ANOVA with Tukey s multiple compariso test agait the DMSO control; F=7.71; degree of freedom=27). In b and c, horizontal lines indicate mean values., not significant.
Supplementary Fig. 9 a 28- IL2RBz 28- IL2RB (FLSL)-z 28- IL2RB-z Control NGFR K562 No cytokine NALM-6 IL-2 K562 No cytokine NALM-6 IL-2 IL-21 PD-L1 (PE) IL-21 TIM-3 (APC-Cy7) PD-1 (Alexa Fluor 488) LAG-3 (PerCP/Cy5.5) 15 1 5 TIM-3 IL-21 Control NGFR IL-2 IL-21 IL-2 Control NGFR none 28- IL2RB-z 28- IL2RB (FLSL)-z 28- IL2RB-z NALM-6 K562 none 2 25 2 15 1 5 28- IL2RB-z 28- IL2RB (FLSL)-z LAG-3 P=.15 4 28- IL2RB-z 6 P=.7 2 1 8 6 4 2 Mean fluorescence inteity PD-L1 PD-1 b Supplementary Fig. 9. Expression of immunoinhibitory molecules in CAR-T cells upon antigen stimulation. The indicated CAR-T cells were cocultured with NALM-6 or K562, or T cells traduced with the control NGFR were treated with IL-2 (3 IU/ml) or IL-21 (5 ng/ml). The expression of PD-1, PD-L1, LAG-3 and TIM-3 was analyzed 24 hours later. Representative FACS plots (a) and the mean fluorescence inteity (b) are shown (n=4; repeated measures one-way ANOVA with Tukey s multiple compariso test for the data for activated CAR-T cells; F=22.8 for PD-1, F=1.52 for PD-L1, F=16.66 for LAG-3, F=2.28 for TIM-3; degree of freedom=19)., not significant. Horizontal lines indicate mean values.
Supplementary Fig. 1 a NALM-6 NALM-6 NALM-6 Cytokine production Day 7 14 21 b Fold expaion of CD8 + CAR-T cells 1 1 4 1 1 3 1 1 2 1 1 1 P=.3 P=.8 P=.2 P=.13 P=.2 P=.7 P=.3 P=.19 P<.1 P=.19 P=.39 P=.33 CAR-T cells Proliferation 1 1 28- IL2RB (FLSL)-z 28- IL2RB-z 28- IL2RB (FLSL)-z 28- IL2RB-z 28- IL2RB (FLSL)-z 28- IL2RB-z c IL-2 production (%) IFN- production (%) 3 2 1 1 8 6 4 2 Day 7 P=.5 P=.7 P=.2 P<.1 Day 14 Day 21 TNF- production (%) 5 4 3 2 1 P=.3 P=.18 IL-2 + IFN- + TNF- + cells (%) 15 1 5 P=.18 28- IL2RB (FLSL)-z 28- IL2RB-z 28- IL2RB (FLSL)-z 28- IL2RB-z P=.13 P=.2 P<.1 28- IL2RB (FLSL)-z P=.12 28- IL2RB-z Stimulation 1st 2nd 3rd Supplementary Fig. 1. Proliferation and cytokine production in the CAR-traduced T cells upon repeated antigen stimulation. (a-c) The CAR-traduced T cells were stimulated weekly with NALM-6 (a). The fold expaion of the CD8 + CAR-T cells at 7, 14 and 21 days after the initial stimulation was calculated (b; n=5 different donor samples, repeated measures one-way ANOVA with Tukey s multiple compariso test; F=26.41 for Day 7, F=52.27 for Day 14, F=83.83 for Day 21; degree of freedom=24). (c) The production of IL-2, IFN- and TNF- was analyzed by intracellular flow cytometry after each stimulation. The frequency of the cells producing individual cytokines and those positive for all cytokines within the CD8 + NGFR + T cell population are shown (n=7 different donor samples, repeated measures one-way ANOVA with Tukey s multiple compariso test for each round of stimulation; F=2.1 (IL-2-1st), 2.57 (IL-2-2nd), 27.4 (IL-2-3rd), 2.54 (IFN- -1st), 2.57 (IFN- -2nd), 2.66 (IFN- -3rd), 6.95 (TNF- -1st), 7.17 (TNF- -2nd), 3.9 (TNF- -3rd), 4.44 (IL-2 + IFN- + TNF- + -1st), 4.87 (IL-2 + IFN- + TNF- + -2nd)), 24.71 (IL-2 + IFN- + TNF- + -3rd); degree of freedom=34)., not significant. Horizontal lines indicate mean values.
Supplementary Fig. 11 a 28- IL2RB (FLSL)-z z Control NGFR PD-1 (Alexa Fluor 488) Mean fluorescence inteity PD-1 PD-L1 LAG-3 TIM-3 P=.4 P=.1 P<.1 28- IL2RB-z 28- IL2RB (FLSL)- z 28- IL2RB-z Control 28- IL2RB-z 28- IL2RB (FLSL)- z 28- IL2RB-z TIM-3 (APC-Cy7) PD-L1 (PE) LAG-3 (PerCP/Cy5.5) b Control 28- IL2RB-z 28- IL2RB (FLSL)- z 28- IL2RB-z Control 28- IL2RB-z 28- IL2RB (FLSL)- z P=.1 28- IL2RB-z Control Supplementary Fig. 11. Expression profiles of exhaustion markers in CAR-T cells after repeated antigen stimulation. (a, b) CAR-T cells were stimulated weekly with NALM-6. Expression profiles of PD-1, PD-L1, LAG-3 and TIM-3 in CD8 + CAR-T cells 7 days after the third stimulation were analyzed by flow cytometry. Representative FACS plots (a) and mean fluorescence inteity (b) are shown (n=5 different donor samples, repeated measures one-way ANOVA with Tukey s multiple compariso test; F=16.17 for PD-1, F=2.97 for PD-L1, F=31.35 for LAG-3, F=56.36 for TIM-3; degree of freedom=29)., not significant. Horizontal lines indicate mean values.
Supplementary Fig. 12 a IL-2-induced genes IL-7-inducd genes IL-15-induced genes P=.178 FDR=.511 P=.649 FDR=.796 P=.444 FDR=.662 P=.313 FDR=.24 P=.143 FDR=.218 P=.59 FDR=.24 b 4 hours z z IL-21-induced genes 4 hours 72 hours 72 hours P=.545 FDR=.656 P=.66 FDR=.676 P<.1 FDR=.49 P<.1 FDR=.11 c 4 hours STAT3 target genes 4 hours 72 hours 72 hours P=.596 FDR=.615 P=.326 FDR=.395 P<.1 FDR=.43 P<.1 FDR=.21 Supplementary Fig. 12. Gene expression profiles of the stimulated CAR-T cells. (a) Gene set enrichment analysis (GSEA) comparing the expression profiles of the z CAR-T cells versus the or CAR-T cells stimulated with NALM-6 for 24 hours (n=4 different donor samples for each group). induced by IL-2, IL-7 or IL-15 treatment were used as the gene sets. (b, c) GSEA of IL-21-induced gene (b) and STAT3 target genes (c) comparing 28- IL2RB-z CAR-T cells and / CAR-T cells stimulated for 4 or 72 hours. A nominal P value for enrichment was calculated by a permutation test, and a false discovery rate (FDR) was calculated to adjust for multiple hypothesis testing.
Supplementary Fig. 13 % tumor cells 1 75 5 25 28- IL2RB (FLSL)- z 28- IL2RB-z 28- IL2RB (FLSL)- z 28- IL2RB-z 28- IL2RB (FLSL)-z 28- IL2RB-z Target cells NALM6-GL K562-CD19 K562 Supplementary Fig. 13. Cytotoxic activity of CAR-T cells. The CAR-traduced T cells were cocultured with the indicated target cells at a ratio of 1:1. The frequency of the residual target cells was determined by flow cytometry (n=4 technical replicates; ordinary one-way ANOVA with Tukey s multiple compariso test for each target cell; F=.48 for NALM6-GL,.49 for K562-CD19,.9 for K562; degree of freedom=19)., not significant. Horizontal lines indicate mean values ± s.d.
Supplementary Fig. 14 Weeks after T cell infusion No T cells z 1 4 6 8 1 Supplementary Fig. 14. Antileukemic effects of the CAR-T cells in vivo. NOD-scid IL2r null (NSG) mice were intravenously injected with the CD19 + leukemia cell line NALM-6 traduced with EGFP-luciferase (NALM6-GL) and were treated with 5x1 6 CAR-T cells 14 days later. In vivo bioluminescent imaging of luciferase activity following the infusion of the CAR-T cells is shown.
Supplementary Fig. 15 a CAR traduction aapc/mokt3 NALM-6 stimulation Day -14-7 CD3 + T cells Day -14 NALM6-GL i.v. b Weeks after T cell infusion 1 4 CAR-T cell i.v. 2 2 c Total photon counts (log1) 9 6 3 2 4 6 8 1 Weeks after T cell infusion No T cell No T cells z 6 8 d Leukemia-related survival (%) g 1 IL-2 (pg/ml) 75 5 25 1 4 8 12 Weeks after leukemia infusion 5 4 3 2 1 P=.1 P=.9 No T cell No T cell IFN- (ng/ml) 15 1 5 e CD8 + CAR-Tcells in the peripheral blood (%) 12 8 4 Days after T cell infusion P=.4 P<.1 P=.8 7 21 Supplementary Fig. 15. Improved therapeutic effects of CAR-T cells by dose escalation. (a) NOD-scid IL2r null (NSG) mice were intravenously infused with the NALM6-GL and treated with two infusio of the 5x1 6 CAR-T cells. (b, c) Leukemia progression was monitored by in vivo bioluminescent imaging of luciferase activity. Individual images (b) and the quantified total photon counts (c) are shown (n=6 mice for each group). (d) Kaplan Meier curve for leukemia-related survival of the mice (n=6 mice for each group, log-rank test). The mice that died of xenogeneic graft-versus-host disease (GVHD) were ceored at the time of death. See Supplementary Table 3 for detailed information. (e) The frequency of CD8 + CAR-T cells in the peripheral blood (n=6; ordinary one-way ANOVA with Tukey s multiple compariso test; F=1. for Day 7, F=3.92 for Day 21; degree of freedom=17). (f) The absolute number of CAR-T cells persisting within the spleen was analyzed at the time of lethal GVHD or on 9 days after leukemia infusion (n=4 for 28z, n=5 for, n=6 for z ; ordinary one-way ANOVA with Tukey s multiple compariso test; F=5.72; degree of freedom=14). The data in d-f are representative of two experiments. (g) Serum was collected from the mice 4 days after the first CAR-T cell infusion, and the concentratio of IL-2, IFN- and TNF- were measured using an enzyme-linked immunosorbent assay (n=6 for each CAR group; ordinary one-way ANOVA with Tukey s multiple compariso test; F=11.16 (IL-2-one), 3.3 (IL-2-two), 2.1 (IFN- -one), 2.57 (IFN- -two), 6.84 (TNF- -one), 3.59 (TNF- -two); degree of freedom=17). IL-2, IFN-, and TNF- were not detected in 8, 12, and 1, respectively, of the 12 No-T cell samples., not significant; nd, not detected. In e-g, horizontal lines indicate mean values ± s.d. P=.3 P=.8 nd No T cell P=.4 TNF- (pg/ml) 1 8 6 4 2 f Absolute number of CAR-Tcells in the spleen P=.9 CAR-T cell infusion One Two One Two One Two P=.3 1 1 8 1 1 6 1 1 4 1 1 2 1 1 P=.3 P=.4 No T cell
Supplementary Fig. 16 a CAR-T cell infusion Day: 5 x 1 6 Day : 5 x 1 6 Day 2: 5 x 1 6 14 12 1 8 6 2 4 6 8 1 14 12 1 8 z 14 12 1 8 6 2 4 6 8 1 14 12 1 8 6 2 4 6 8 1 * * * * 6 6 6 2 4 6 8 1 2 4 6 8 1 2 4 6 8 1 Weeks after T cell infusion Weeks after T cell infusion Weeks after T cell infusion 14 12 1 8 14 12 1 8 b NGFR (Pacific Blue) 1.7 25.8 16.3 12. 21.6 29.3 CD45 (APC) Supplementary Fig. 16. The development of xenogeneic graft-versus-host disease (GVHD) by CAR-T cells after leukemia eradication. (a) Serial weight monitoring of NALM-6-bearing mice infused with the indicated number of CAR-T cells (n=6 mice for each group). Relative body weight compared with the weights prior to T cell infusion are shown. The development of lethal GVHD is denoted by *. (b) Persistence of CAR-T cells within the spleen of the mice that received two infusio of 5x1 6 CAR-T cells was analyzed at the time of lethal GVHD or at 9 days after leukemia infusion. Representative FACS plots analyzing CD45 + NGFR +/- cells within the spleen of 6 mice for each group are shown.
Supplementary Fig. 17 a Infusion 5 million 1 million % body weight 14 12 1 8 6 2 4 6 8 1 14 12 1 8 z 14 14 12 12 1 1 6 2 4 6 8 1 Weeks after T cell infusion % body weight * 8 ** * * * * * 6 2 4 6 8 1 % body weight 8 14 14 12 12 1 1 * * * * * * 6 2 4 6 8 1 8 8 * * * * * * * * 6 6 2 4 6 8 1 2 4 6 8 1 Weeks after T cell infusion Weeks after T cell infusion * * * * * b Overall survival (%) c 5 million 1 million 1 8 6 4 2 2 4 6 8 1 Absolute number of CAR-Tcells in the spleen Weeks after T cell infusion 1.5 1 6 1. 1 6 5. 1 5. Overall survival (%) z Supplementary Fig. 17. The development of xenogeneic graft-versus-host disease by CAR-T cells in the absence of antigen. Tumor-free NSG mice were irradiated with 1.5 Gy, and traplanted with 5 or 1 million of CAR-T cells. (a) Serial weight monitoring of the mice. Relative body weights compared with the weights prior to T cell infusion are shown. The development of lethal GVHD is denoted by * (n=6 mice for each group). (b) Overall survival of the mice traplanted with CAR-T cells (n=6 mice for each group, log-rank test). (c) Tumor-free NSG mice were irradiated with 1.5 Gy and adoptively traferred with 5x1 6 CAR-T cells. The mice were sacrificed 14 days after T cell infusion, and the absolute number of the CAR-T cells within the spleen was analyzed (n=6 mice; ordinary one-way ANOVA with Tukey s multiple compariso test; F=2.54; degree of freedom=17). Representative of two experiments., not significant. Horizontal lines indicate mean values ± s.d.
Supplementary Fig. 18 a No T cells 28- IL2RB-z SSC Debris exclusion FSC FSC-W Doublets exclusion SSC-W FSC-H CD19 (PE) SSC-H HLA-A2 (FITC) 1 * * 1.1 Day 7.1 28- IL2RB-z CD45 (APC) NGFR (Pacific Blue) b CD8+ CAR-Tcells in the peripheral blood (%) c Time of tumor progression Supplementary Fig. 18. Relapse of the melanoma cell line A375-CD19 after treatment with CAR-traduced T cells. (a, b) NSG mice were subcutaneously injected with A375 melanoma cells traduced with CD19 (A375-CD19) (day -21) and infused with 5x15 CAR-T cells on day and 4. The mice were sacrificed when the tumor size exceeded 3 mm3. (a) Progressing tumor cells were analyzed for CD19 expression. Cells were co-stained with HLA-A2 to discriminate the A375CD19 tumor cells. (b) Persistence of the CAR-T cells within the tumor was analyzed when the tumor size exceeded 3 mm3. In a and b, representative FACS plots from seven mice in each group are shown. (c) The frequency of CD8+ CAR-T cells in the peripheral blood 7 days after CAR-T cell infusion or at the time the tumor mass exceeded 3 mm3 (n=7 mice; ordinary one-way ANOVA with Tukey s multiple compariso test for each time point; F=4.19 for Day 7, F=.71 for Time of tumor progression; degree of freedom=2). * P<.1., not significant. Horizontal lines indicate mean values ± s.d.
Supplementary Table 1. Amino acid sequencing of the CAR signaling domai used in this study. IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLAC YSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFA AYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEM GGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLS TATKDTYDALHMQALPPR Blue: CD28 Black: CD3z 28-IL2RB-z z 28- IL2RB (FLSL)-z z TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPL AGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEE Black: CD8 EEGGCELPRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGR Green: 4-1BB DPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLY Black: CD3z QGLSTATKDTYDALHMQALPPR IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLAC YSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFA AYRSNCRNTGPWLKKVLKCNTPDPSKFFSQLSSEHGGDVQKWLSSPFPS SSFSPGGLAPEISPLEVLERDKVTQLLLQQDKVPEPASLSSNHSLTSCFTNQ Blue: CD28 GYFFFHLPDALEIEACQVYFTYDPYSEEDPDEGVAGAPTGSSPQPLQPLSG Red: IL2RB EDDAYCTFPSRDDLLLFSPSLLGGPSPPSTAPGGSGAGEERMPPSLQERV Black: CD3z with PRDWDPQPLGPPTPGVPDLVDFQPPPELVLREAGEEVPDAGPREGVSFP YXXQ WSRPPGQGEFRALNARLPLNTDAYLSLQELQGQDPTHLVRVKFSRSADAP AYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLY NELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDAYRHQA LPPR IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLAC YSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFA Blue: CD28 AYRSNCRNTGPWLKKVLKCNTPDPSKFFSQLSSEHGGDVQKWLSSPFPS Red: IL2RB with SSFSPGGLAPEISPLEVLERDKVTQLLPLNTDAYLSLQELQGQDPTHLVRVK internal deletion FSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRK Black: CD3z with NPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTY YXXQ DAYRHQALPPR IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLAC Blue: CD28 YSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFA Red: IL2RB with AYRSNCRNTGPWLKKVLKCNTPDPSKFFSQLSSEHGGDVQKWLSSPFPS internal deletion SSFSPGGLAPEISPLEVLERDKVTQLLPLNTDAFLSLQELQGQDPTHLVRVK and FLSL FSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRK Black: CD3z with NPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTY YXXQ DAYRHQALPPR IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLAC YSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFA Blue: CD28 AYRSNCRNTGPWLKKVLKCNTPDPSKFFSQLSSEHGGDVQKWLSSPFPS Red: IL2RB with SSFSPGGLAPEISPLEVLERDKVTQLLPLNTDAYLSLQELQGQDPTHLVRVK internal deletion FSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRK Black: CD3z NPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTY DALHMQALPPR
Supplementary Table 2. Differentially expressed genes in different CAR-T cells at the indicated time points (P<.1 with repeated measures one-way ANOVA, and fold change >1.5 or <-1.5). IL-21-induced genes, STAT3 target genes and genes included in both gene sets are highlighted in red, blue and green, respectively. 4 hours after NALM-6 stimulaiton (356 genes) BBz vs 28z vs BBz BBz z vs BBz BBz z vs BBz BATF3.15 1.3 1.43 1.86 CLECL1.9 1.41 1.24 1.75 ABCB1.87 1.5-1.15 1.31 TWSG1.61 1.5 1.5 1.58 KCNA3.15 1.8 1.43 1.55 IL31.51 2.73-1.58 1.72 MFSD2A.3 1.73 1.4 1.8 MIR548C.94 1.72 1.9 1.87 CTDSP2.93-1.75 1.4-1.26 PRDM1.46 1.32 1.17 1.54 FIGNL1.27-1.4 1.51 1.45 CATSPERB.5 1.93-1.33 1.45 NKG7.34 1.27 1.18 1.5 KAT2B.52 1.51 1.17 1.76 WRAP53.44-1.52 1.28-1.18 CTLA4.3 2.9 1.75 3.65 ELL2.55 1.43 1.2 1.72 SKA1.19-1.96 1.44-1.36 SEC61A2.14 1.5 1.75 1.83 POFUT1.14 1.31 1.26 1.65 TTK.66-1.6 1.3-1.23 METRNL.63 1.29 1.44 1.86 ST8SIA4.2 1.31 1.26 1.65 LINC892.3 1.61-1.24 1.3 GPR132. -1.5 1.69 1.6 ADAM8.14 1.9 1.39 1.51 FSCN1.91-1.6 1.29-1.24 ABTB2.1 1.77 1.12 1.99 INPP4B.6 1.34 1.24 1.66 RMI1.6-1.86 1.39-1.34 CYTIP.9 1.7 1.4 1.5 OAS2.18-1.27 1.61 1.26 ALG1.56-1.69 1.32-1.28 JAK3.58-1.19 1.54 1.29 MIR1471.68-1.74 1.82 1.4 GOLGA8DP.54 1.61-1.25 1.29 MLLT6.7 1.2 1.27 1.53 MYO1G.29-1.2 1.55 1.29 IL2.67 1.65-1.27 1.3 RORA.11 1.29 1.2 1.56 PIK3R5.3 1.45 1.15 1.67 AGFG2.11-1.5 1.23-1.21 BATF.69 1.32 1.42 1.87 ANTXR2.2 1.29 1.22 1.57 NUDT6.43-1.53 1.24-1.24 ICOS.24 1.7 1.48 1.58 HAPLN3.21-1.18 1.51 1.28 HLA-DRA.44-2.65 1.61-1.65 KIF3A.34-1.42 1.69 1.19 MT1CP.53-1.2 1.52 1.27 CHEK1.11-1.57 1.23-1.27 IL5.5 2.3 1.85 3.76 SEC24D.6 1.17 1.28 1.5 CD74.28-1.53 1.21-1.27 LOC11928173.6 2.7 1.61 3.33 MDFIC.8 1.38 1.17 1.6 FAM9A1P.47 1.53-1.27 1.21 SH3BP5.23 1.22 2.12 2.58 PIM2.4 1.42 1.14 1.63 PRIM1.76-1.68 1.26-1.33 MAF. 1.41 1.8 2.54 SEC31B.5 1.26 1.22 1.54 PTTG3P.33 1.54-1.29 1.2 APOL1.8 1.4 1.8 2.51 CD83. 1.51 1.9 1.65 CBX5.8-1.52 1.18-1.28 CD86.3 1.59 1.62 2.56 MX1.79 1.23 1.23 1.51 HLA-DPA1.98-1.54 1.19-1.3 CCR4.2-1.16 2.16 1.87 LOC11929531.16-1.25 1.51 1.21 C7orf31.58-1.66 1.23-1.35 MSC.5 1.53 1.58 2.42 SLC7A11.78 1.23 1.22 1.5 GPR87.6 2.66-1.7 1.57 FAM46C.3 1.29 1.73 2.24 SGPL1.11 1.42 1.11 1.57 RFC4.39-1.8 1.27-1.41 RCAN2.54 1.44 1.61 2.32 ACOX3.19 1.45 1.9 1.58 CENPU.3-1.5 1.16-1.3 IL22.1 1.63 1.45 2.36 TRPS1.71 1.31 1.16 1.51 IL1R1.67 1.51-1.3 1.16 APOL2.16 1.44 1.56 2.24 LOC15373723.17 1.53 1.5 1.61 DHCR24.19-1.79 1.26-1.42 SYTL3.4 1.35 1.61 2.17 CST7.54 1.31 1.15 1.51 NCAPH.7-1.74 1.24-1.4 TOX.27 1.22 1.71 2.7 LOC34156.97-1.41 1.55 1.1 CTTNBP2NL.45-1.61 1.19-1.35 GZMH. 1.58 1.46 2.31 MAML2.46 1.43 1.9 1.56 ANLN.13-1.73 1.22-1.41 DDX6.14 1.4 1.82 1.9 LMNB1.27-1.34 1.52 1.13 E2F1.72-1.67 1.18-1.41 C1orf128.8 1.82 1.29 2.35 TPI1P3.4 1.32 1.14 1.5 FOXM1.43-1.5 1.11-1.35 TNFRSF8.24 1.4 1.77 1.85 ADRB2.63 1.4 1.1 1.53 VTRNA1-2.8 1.69-1.43 1.18 FCRL3.13 1.45 1.47 2.12 LOC11929536.23-1.38 1.52 1.1 MIR4773-2.15 1.56-1.39 1.13 NUGGC.11 1.2 1.74 1.78 SLC41A2.28 1.46 1.6 1.55 BRIP1.61-1.56 1.12-1.39 IL6R.23 1.3 1.52 1.98 UBXN1.91-1.39 1.5 1.8 ITGAV.69 1.58-1.4 1.13 LY96.21 1.19 1.58 1.88 SOD2.44 1.47 1.3 1.52 MTX3.2 1.83-1.5 1.21 LAMP3.4 1.48 1.39 2.7 LOC15376626. 2.9-1.4 2.78 TPX2.6-1.75 1.18-1.48 INSIG1. 1.62 1.32 2.13 ITGA2.13 2.12-1.4 2.4 LGALS9B.39-1.57 1.11-1.42 HERC6.62-1.5 1.72 1.65 MCOLN2.29 2.4-1.3 1.98 GTSE1.64-1.99 1.26-1.58 PDE4DIP.54 1.26 1.49 1.88 LOC15372752.46 2.41-1.21 2 HIST1H3G.43-1.79 1.19-1.51 PROK2.3 2.14 1.6 2.28 FNDC9.23 1.73-1.2 1.71 ECT2.1-1.72 1.16-1.48 FGFBP2.13 1.5 1.31 1.97 MIR222.21 2.11-1.15 1.83 GCNT1.21 1.98-1.58 1.25 LTA.1 1.31 1.42 1.86 IL4.43 1.82-1.5 1.73 MIR431.31-1.66 1.13-1.47 ADGRG1.19 1.9 1.54 1.68 KLF3.62 1.52-1.1 1.51 NDC8.69-1.66 1.13-1.47 STRIP2.15 1.99 1.7 2.12 TGFBR3.98 1.63-1.6 1.55 CDC2.9-1.64 1.12-1.47 F5.38 1.59 1.23 1.96 TNF.2 1.8-1.15 1.56 CCL2.1 2.45-1.75 1.4 GALNT3. 1.37 1.34 1.84 NIPA1.14 1.8-1.17 1.54 PLK1.23-1.86 1.2-1.55 GSAP.37 1.31 1.37 1.79 PLEK.8 2.13-1.29 1.66 CKAP2.14-1.58 1.9-1.45 TRGV2.82 1.21 1.42 1.73 B3GNT5.1 1.57-1.8 1.45 KIAA1841.74-1.96 1.23-1.59 EOMES.11 1.18 1.44 1.7 LOC1537952.88 1.61-1.1 1.47 BIRC5.3-1.76 1.15-1.53 RASA2.25 1.15 1.46 1.68 LINC861.76 1.61-1.11 1.44 NUSAP1.9-1.71 1.12-1.52 LIMA1.12 1.17 1.44 1.69 CCL3L3.7 1.71-1.16 1.47 SCCPDH.45-1.55 1.6-1.46 TMEM173.71-1.12 1.65 1.48 PIGV.41 1.53-1.1 1.39 UBE2C.1-1.5 1.4-1.45 MIR4326.72 1.96 1.5 2.7 ZNF28D.93-1.54 1.39-1.11 ACAT2.94-1.51 1.3-1.47 MIR518A1.37-1.8 1.62 1.5 CDC42EP3.36 1.85-1.24 1.5 DTL.29-1.54 1.4-1.48 LOC1192792.5 1.21 1.41 1.71 MIR96.78 1.63-1.16 1.4 CENPE.4-1.68 1.9-1.54 CD274.2 1.39 1.28 1.78 BCL9L.82-1.63 1.4-1.17 MTBP.9-1.64 1.7-1.54 EPAS1.19 1.28 1.34 1.71 PTPN6.43-1.69 1.42-1.19 NCAPG2.4-1.59 1.5-1.52 PFKFB3.12 1.14 1.41 1.61 DLGAP1.54 1.54-1.14 1.35 GINS1.88-1.52 1.2-1.49 SETBP1.71 1.32 1.3 1.72 LINC944.57 1.74-1.22 1.42 ADRBK1.17-1.51 1.1-1.5 ST6GAL1.3-1.2 1.65 1.37 MYCBP.16-1.5 1.31-1.14 CCNB1.64-1.5 1-1.49
vs 28z vs BBz z vs BBz z vs BBz KIF11.3-1.86 1.14-1.64 ADRBK2.4-1.19-1.31-1.55 RASGEF1B.67 1.27-1.93-1.52 PRC1.3-1.55 1.2-1.53 CD9.59-1.8-1.2-1.85 ANKRD3B.84-1.73-1.27-2.19 SQLE.4-1.56 1.2-1.53 SMOX.1 1.84-1.86-1.1 PIP5K1B.75-1.86-1.23-2.29 RAPH1.6 1.85-1.66 1.12 ABHD2.33 1.8-1.5-1.38 BTLA.1 1.6-1.85-1.73 UBE2T. -1.9 1.13-1.68 FGFR1.2-1.29-1.26-1.62 CD2.8 1.3-1.86-1.82 ATAD2.2-1.71 1.6-1.62 IKZF2.46 1.9-1.5-1.38 CD72.16 1.24-2.4-1.65 INSL6.28 1.63-1.59 1.2 SERPINE2.15 1.15-1.54-1.34 MTSS1.6-1.88-1.28-2.41 FANCI.3-1.74 1.6-1.64 SLAMF6.8 1.14-1.53-1.35 TREML5P.16-1.22-1.67-2.4 MKI67.52-1.71 1.5-1.63 TRAV8-4.53-1.18-1.32-1.56 NR4A2. -1.3-1.84-1.88 USP17L9P.44 1.76-1.66 1.6 FAM213A.3-1.24-1.29-1.6 INSM1.18-1.1-1.79-1.97 DLGAP5. -1.72 1.4-1.65 LOC15376878.1-1.13-1.36-1.53 FGL2.7 1.9-1.97-1.8 TICRR.2-1.84 1.7-1.71 ZG16B.24-1.46-1.18-1.72 ID3.1-1.35-1.61-2.18 CCNA2. -2.6 1.15-1.79 INPP5F.11-1.62-1.11-1.79 PHEX.9-1.76-1.47-2.59 TOP2A.3-1.7 1.1-1.69 GREB1.94-1.23-1.3-1.6 VCAM1.68-2.31-1.24-2.86 YPEL3.18-1.81 1.3-1.75 TSPAN2.4-1.78-1.5-1.86 ARMC9.1-1.54-1.63-2.5 FAM111B.3-2.11 1.13-1.87 CD5.11-1.42-1.21-1.71 FAM72A.21 1.19-2.25-1.89 COL6A2.3-2.27 1.17-1.94 FLNB.31-1.15-1.36-1.56 ARHGAP42.2 1.27-2.34-1.83 CCNB2.68-1.83 1.2-1.8 HHLA3.9-1.5-1.43-1.5 IGLC7.89-2.23-1.29-2.89 HLA-DQA2.34-2.82 1.33-2.12 ARHGAP31.2-1.56-1.15-1.79 NCEH1.6 1.1-2.24-2.4 EIF1AX.6 1.84-1.82 1.1 DUSP18.9 1.58-1.8-1.14 COL6A1.1-2.43-1.28-3.1 TNFSF4.13-2.76 1.27-2.17 MYO1E.82-1.13-1.38-1.56 KIAA1324.11-2.81-1.18-3.32 NCAPG.2-2.8 1.5-1.99 DGKA.26-1.46-1.2-1.75 CCL1.9-1.27-2.5-2.6 CCR8.33-3.43 1.3-3.33 UPRT.58-1.6-1.44-1.52 DHRS3. -1.54-1.93-2.96 CDK1.47-1.49-1.1-1.5 KIT.36-1.83-1.5-1.92 NHS. -1.88-1.91-3.59 NCAPD3.11-1.54-1.1-1.55 RSPH4A.23-1.4-1.46-1.51 CD16. 1.51-3.33-2.21 NEK2.35-1.52-1.2-1.55 SGK223.1 1.25-1.65-1.32 SERPINE1.1-1.54-2.2-3.38 THYN1.8-1.42-1.6-1.51 SNORD113-9.76-1.43-1.22-1.75 ISM1. 1.13-3.8-2.72 MCM1.41-1.55-1.1-1.57 EVI2A.27 1.98-1.99-1.1 CCL22.1-2.96-3.42-1.13 LOC15378541.54 1.45-1.53-1.5 GJB2.26-1.18-1.38-1.63 IL23A.11 1.49-1.57-1.5 HSD17B11.63-1.1-1.5-1.52 ASPM.1-1.5-1.5-1.58 CCND1.49 1.16-1.62-1.4 PDCD4.31-1.41-1.9-1.54 DOCK5.5-1.42-1.25-1.77 FLJ44385.1 1.31-1.5-1.14 MYADM.2-1.25-1.34-1.68 CCP11.8-1.3-1.15-1.5 LOC15379311.86 1.41-1.77-1.26 HSPB1.49-1.56-1.4-1.61 RILPL2.4 1.15-1.62-1.41 LEF1.89-1.42-1.1-1.56 TLE4.36-1.2-1.5-1.54 CAMK2D.6 1.34-1.54-1.14 CD84.13 1.22-1.67-1.37 P2RY1.49-1.27-1.18-1.51 CENPH.54-1.3-1.32-1.72 SPTBN1.55-1.35-1.14-1.55 EXO1.8-1.54-1.2-1.85 CCDC141.21-1.49-1.8-1.61 LOC11927482.91 1.52-1.84-1.21 DRAM2.15-1.31-1.17-1.53 NELL2. -1.23-1.37-1.68 RRM1.1-1.62-1.3-1.68 ENOX1.81-1.61-1.17-1.89 E2F8.44-1.3-1.18-1.53 KIF23. -1.94-1.4-2.3 ASB2.71-1.43-1.12-1.6 SPRY1.9 1.13-1.63-1.44 BNIP3L.1-1.42-1.12-1.6 TMEM14C.79-1.5-1.5-1.57 HMGB2. -1.57-1.6-1.67 XCL2.3 1.3-1.56-1.51 PRR11.7-1.56-1.7-1.66 PIKFYVE.17-1.18-1.41-1.67 C12orf57.7-1.4-1.14-1.59 PTPN3.25-1.13-1.45-1.65 CHST1.94 1.28-1.53-1.2 TESK1.3 1.1-1.63-1.48 TBC1D2.8-1.25-1.22-1.51 HVCN1.11-1.74-1.14-1.98 AFF3.7-1.45-1.12-1.62 RALGAPA2.6 1.1-1.58-1.56 CXorf21.7-1.45-1.12-1.62 FCMR.15 1.24-1.75-1.41 FAT1.37-1.28-1.21-1.54 SLC35E4.16 1.31-1.79-1.37 LCK.2-1.23-1.23-1.52 LAYN.4-1.74-1.16-2.1 MIR3646.7-1.48-1.11-1.64 IL3.2 1.19-1.72-1.45 TRAF5.4 1.2-1.5-1.25 SESTD1.81-1.17-1.46-1.71 CENPF.6-1.54-1.9-1.67 LOC1576.61-1.5-1.27-1.91 HMMR. -1.66-1.4-1.72 TCF7.2-1.44-1.3-1.88 POLR3C.2 1.19-1.5-1.26 GPR155.19 1.32-1.83-1.38 MELK.83-1.46-1.13-1.65 NLRP6.24 1.6-1.68-1.58 SMC2.29-1.53-1.1-1.68 FGF2.68-1.21-1.48-1.79 CENPV.51-1.21-1.26-1.52 PLXNC1.1 1.7-1.72-1.6 SUOX.23-1.6-1.7-1.72 ZCWPW2.25-1.93-1.15-2.21 TYMS.12-1.61-1.7-1.72 FAM12A.12-1.21-1.52-1.84 NT5E.45 1.56-1.72-1.1 MGAT5.7 1.18-1.82-1.54 GPATCH11.4-1.33-1.21-1.61 CD27. -1.7-1.63-1.76 ARMCX6.64-1.14-1.32-1.51 PRCP. -1.5-1.67-1.75 CDKN3.83-1.23-1.27-1.57 JAG1.16-1.48-1.38-2.5
24 hours after NALM-6 stimulaiton (429 genes) vs 28z vs BBz vs 28z vs BBz vs 28z vs BBz IL1.15 1.18 5.77 6.83 MIR58.76-1.8 1.89 1.75 LOC72988.4 1.54 1.14 1.76 IL1R1.12 1.48 1.59 2.36 CD274.5 1.44 1.49 2.14 PCCA.2-1.8 1.51 1.39 KLRC1.14-1.4 2 1.92 SLC2A3.14-1.9 1.88 1.73 MTM1.14-1.51 1.74 1.15 F2R.2-1.1 2.3 1.85 LOC15376145.28 1.1 1.72 1.89 SETBP1.2 1.53 1.14 1.74 GZMB.42 1.4 1.43 1.99 PALLD.36-1.6 1.81 1.71 CASP8.46-1.17 1.55 1.32 PRDM1.9 1.41 1.31 1.85 AIM2.6-1.9 1.83 1.68 FCMR.78 1.24 1.28 1.59 CCR5.94-1.27 1.76 1.38 TNIP3.8-1.7 1.82 1.69 PWARSN.56 1.26 1.27 1.59 IL1R2.5 1.27 1.35 1.71 LOC389765.55 1.51 1.39 2.9 CLK1.61-1.19 1.55 1.3 KSR1.9-1.12 1.57 1.4 ABCA2.4 1.7 1.68 1.79 RASGRF2.89 1.13 1.34 1.51 GZMA.33-1.52 1.76 1.16 RGS9.2-1.18 1.87 1.59 PGAM1.5 1.38 1.2 1.65 NKG7.36 1.51 1.13 1.7 ANKRD2A5P.52 1.53 1.36 2.8 LAG3.27 1.78 1.2 1.81 GCNT1.17 1.25 1.21 1.5 TMED1P1.49 1.6 1.67 1.77 GLUD1.37 1.14 1.32 1.51 BMPR1A.32 1.51 1.2 1.54 MIR3685.16 1.52 1.36 2.7 PBXIP1.76-1.39 1.64 1.18 IL21.38-1.12 2.87 2.56 XRRA1.5 1.6 1.65 1.75 CIITA.66-1.3 1.59 1.22 ABLIM1.98-1.2 2.2 2.15 ACVR1.3 1.17 1.56 1.83 LOC22729.88-1.14 1.5 1.31 GALNT1.17-1.2 2.9 2.4 TC2N.2-1.68 2.11 1.26 CST7.9 1.45 1.15 1.66 CTLA4.2 1.2 1.98 2.2 ARRDC3. -1.25 1.86 1.49 TMEM14EP.73-1.34 1.61 1.19 SBNO2.15 1.2 1.79 1.82 MIR3945.52 1.25 1.49 1.86 MAP3K5. -1.35 1.6 1.19 CD28.6-1.22 1.96 1.61 TANC2.17 1.2 1.51 1.82 TMEM71.43-1.46 1.66 1.13 SPEF2.7 1.18 1.6 1.88 LINC-PINT.25 1.28 1.46 1.86 NIPAL4.39 1.41 1.15 1.63 MFAP3.25-1.11 1.79 1.61 BCAT1.43 1.4 1.62 1.69 SERPINI1.37-1.45 1.64 1.13 SOX5.3 1.2 1.64 1.68 RTKN2. -1.28 1.85 1.44 LGALS3.78 1.28 1.21 1.55 NFIA.4 1.14 1.5 1.72 RNF213.5-1.48 1.95 1.32 LOC15376177.5 1.31 1.19 1.56 ITGA3.56-1 1.52 1.52 PGAP1.5 1.6 1.58 1.67 CCL4L1.71 1.68 1.2 1.71 RFX3.42-1.41 1.54 1.9 LOC11638.72-1.37 1.88 1.37 ZNF11.6-1.24 1.51 1.22 BCL3.2-1.18 2.9 1.77 KLRAP1.4 1.2 1.59 1.62 SNORD127.38 1.26 1.21 1.52 BATF.3-1.3 1.75 1.7 SNORD11B.33 1.25 1.42 1.78 VAMP4.24-1.28 1.52 1.19 SOCS3.3 1.34 1.43 1.92 CD226.7 1.4 1.32 1.86 MYO6.4 1.46 1.1 1.61 FOSL2.2 1.25 1.34 1.67 TMEM65.4 1.3 1.56 1.61 SLC7A5P2.88 1.49 1.9 1.62 MCTP2.3-1.11 1.57 1.41 SIK3-IT1.2 1.42 1.31 1.86 FKBP14.79 1.27 1.18 1.51 GPR18.25-1.58 1.66 1.5 MIR573.34-1.16 1.7 1.47 CLU.45-1.37 1.55 1.13 OSTF1.1-1.38 1.51 1.9 IL1RAP.99 1.14 1.48 1.69 C1orf128.69 1.45 1.8 1.56 TNFRSF8.11 1.34 5.99 8.5 ZC3H12D.61-1.6 1.62 1.53 SLC25A4.82 1.63 1 1.64 LOC15373955.22 2 3.58 7.16 NUGGC.52 1.13 1.48 1.67 ZNF449.8-1.34 1.5 1.12 IL26.9-1 4.42 4.41 IGANRP.16-1.7 1.61 1.51 ARHGEF35.17 1.51 1.4 1.57 EPAS1.45 2 2.87 5.74 SERPINB1.37-1.5 1.6 1.52 LOC153795.5-1.46 1.54 1.5 COL6A3.49 1.8 3.41 3.69 LOC154761.52 1.11 1.48 1.64 ADAMTS6.6 1.44 1.4 1.5 RCAN2.4 1.2 3.2 3.9 CCR1.4-1.55 1.89 1.22 ADCY3.79 1.44 1.4 1.5 LOC11928173.11 1.7 2.17 3.69 ADD3.57-1.46 1.84 1.26 F5. 2.75-1.22 2.25 TRERF1. -1.39 3.29 2.36 HLA-DRA.52-1.64 1.92 1.18 EMP1.86 2.37-1.13 2.9 SLCO4C1.84-1.15 3.1 2.61 TXK.74-1.23 1.7 1.38 TP53INP1.45-1.92 1.84-1.4 IL7R.23-1.66 3.24 1.95 NARF.16 1.1 1.53 1.54 MYOF.7 1.93-1.1 1.76 GLUL.43 1.13 2.39 2.71 FAM184A.18 1.3 1.51 1.56 BNIP3.96 1.53-1.1 1.51 ZNRF1.3 1.9 2.42 2.64 RRAGC.5 1.19 1.4 1.67 ALOX5AP.1-1.69 1.55-1.9 ZBED2.57 1.12 2.33 2.6 RPRD1A.5-1.13 1.63 1.44 LOC729461.13-1.58 1.5-1.5 ANKRD2A1.26 1.17 2.12 2.48 PARP9.1-1.3 1.73 1.33 HERC6.31-1.73 1.53-1.13 SULT1B1.22-1.14 2.37 2.8 KLRD1.78 1.5 1.49 1.56 GZMK.51-2.74 1.86-1.47 DDIT4.25-1.14 2.27 1.99 ATP2B4.8-1.73 1.92 1.11 SIRPB1.23-1.73 1.51-1.15 SATB1.1-1.15 2.24 1.95 CRYZ.2-1.14 1.61 1.42 PAM. 1.91-1.22 1.56 STRIP2.4 1.34 1.79 2.4 SNORD99.11 1.88 1.5 1.97 GPSM2.32-1.69 1.44-1.18 SH3BP5.9-1.2 2.27 1.9 HNRNPU.55 1.1 1.5 1.51 RHOU.55-1.5 1.36-1.1 FAM46C.3-1.28 2.34 1.82 LINC484.35 1.36 1.28 1.73 SAMD9L.11-1.82 1.48-1.23 GIMAP7.1-2.15 2.82 1.31 GAB3. -1.24 1.66 1.34 ENPP4.9 1.5-1.11 1.34 GPR16.8 1.24 1.83 2.27 SPATS2L.11-1.3 1.52 1.47 ERP27.9-1.77 1.44-1.23 GIMAP4.5-2.4 2.67 1.31 TRIB2.74-1.66 1.86 1.12 VDR.12 1.55-1.15 1.35 PRKAR2B.33 1.24 1.76 2.19 LOC1996286.8-1.9 1.95 1.3 EVI2B.9-1.85 1.46-1.27 FURIN.1-1.26 2.17 1.73 TCAF2.3-1.2 1.5 1.48 MVB12B.7-1.54 1.33-1.16 STOM.4-1.18 2.11 1.78 ARID3B.56 1.27 1.31 1.66 CNN2.44-1.51 1.31-1.16 KDSR.17 1.24 1.73 2.15 PLGLA.88 1.21 1.34 1.61 TJP2.4 1.86-1.29 1.44 SLC27A2.3 1.3 1.91 1.96 SLC9A9. -1.72 1.86 1.8 GS2.33 1.9-1.32 1.45 GPR183.6-1.5 1.97 1.87 RNF157.4 1.16 1.36 1.58 SNORD35B.41 1.72-1.25 1.38 MAF.14 1.87 1.33 2.48 GALNT3.2 1.11 1.39 1.55 DKFZP586I142.36-1.68 1.35-1.24 LUCAT1.3-1.24 2.1 1.68 CYP4F35P.9 1.7 1.42 1.52 DYNC2LI1.7 1.64-1.23 1.33 PIM2.3 1.56 1.44 2.23 SLC39A1.2-1.22 1.6 1.32 SLFN5.42-2.7 1.49-1.39 FNIP2.7 1.23 1.64 2.2 LOC11927841.43 1.46 1.19 1.73 SAMD9.44-1.54 1.29-1.2 GZMH. 1.62 1.4 2.26 GPR87.14 1.6 1.12 1.78 LOC1537435.66-1.59 1.3-1.22
vs 28z vs BBz vs 28z vs BBz vs 28z vs BBz CTH.535 1.55-1.23 1.26 CYLD.29-1.24-1.24-1.54 ETS2.16-1.3-1.38-1.8 CINP.9847 1.66-1.29 1.29 KIAA922.32-1.19-1.27-1.51 TNFSF14. 1.52-1.93-1.26 FAM65B.433-1.66 1.28-1.29 OR51A2.35-1.19-1.27-1.51 FAM69A. 1.22-1.76-1.44 SMAD3.144-1.54 1.23-1.25 NFIX.72 1.18-1.51-1.28 FOXP1.81 1.21-1.75-1.45 NCKAP1.3843 1.8-1.36 1.33 TNFAIP3.1 1.58-1.72-1.8 FUT8.55 1.1-1.61-1.6 KIF14.2568-1.52 1.21-1.26 PDE7A.23-1.33-1.2-1.6 SERTAD2.3-1.19-1.47-1.74 SNORA71A.693 1.57-1.29 1.22 UBASH3B.1 1.38-1.63-1.18 LOC15377447.5 1.4-1.64-1.58 GVINP1.4866-1.5 1.18-1.28 SNTB1.8-1.2-1.28-1.53 NFE2L3.1 1.35-1.86-1.37 GNPDA1.47 2.3-1.48 1.37 CLIC4.7 1.27-1.57-1.24 INPP5F.69-1.15-1.51-1.73 APOBEC3H.4123-1.56 1.19-1.31 LOC15373187.91-1.56-1.1-1.72 PDCD4.48-1.16-1.5-1.75 CCNG2.279-1.57 1.19-1.32 UNQ6494.71-1.22-1.27-1.55 ARHGAP31.39 1.9-1.7-1.56 LRMP.7492-1.62 1.2-1.35 HIVEP3.45 1.38-1.64-1.19 P2RY14.4-1.78-1.18-2.11 CCR2.834-2.26 1.42-1.59 RAB9A.3-1.14-1.33-1.51 CRIM1.9 2.1-2.25-1.7 IQCF3.2961 1.68-1.41 1.19 LAX1.4 1.24-1.57-1.27 ATXN1.35-1.2-1.66-1.69 NFATC1.9257 1.51-1.35 1.12 CD72.62 1.58-1.75-1.11 SYPL1.3-1.29-1.46-1.89 TRGJP1.89-1.62 1.15-1.4 LOC15379695.87 1.62-1.77-1.9 AMICA1.1 1.33-1.92-1.45 KCCAT198.74-2.35 1.41-1.67 LITAF. 1.28-1.61-1.25 NEK6.5 1.5-1.73-1.65 GLIPR1.2-1.56 1.9-1.42 IL1RA.12-1.16-1.32-1.54 TLR5.11-1.93-1.16-2.24 RGCC.2 1.54-1.42 1.8 XIRP1.99 1.81-1.85-1.2 TNFSF11.21 1.1-1.71-1.7 LOC11929929.37 1.53-1.42 1.8 SNX11.5 1.11-1.51-1.36 MAL.51 1.6-1.75-1.66 MCOLN2.49 1.52-1.44 1.6 KIAA1147.57 1.19-1.56-1.32 PSTPIP2.1 1.99-2.28-1.14 MYBL1.94-1.55 1.5-1.48 MIR1249.59-1.45-1.18-1.71 IKZF2.7-1.6-1.32-2.1 TNFSF13B.62-1.5 1.2-1.46 DNAJC5B.61-1.81-1.3-1.86 COL6A1.12-1.27-1.51-1.93 SIRPG.6-1.86 1.16-1.6 ARHGEF3.22-1.11-1.37-1.52 SNX9.3 1.3-1.95-1.5 RHOB. 1.71-1.55 1.1 OAS3.81-1.3-1.26-1.64 ISM1.44 1.35-1.99-1.48 MIR155HG.15 1.72-1.57 1.9 KRT7.79-1.32-1.25-1.66 NCF2.11 1.4-1.77-1.71 GPR15.5-1.86 1.12-1.66 RAB3.93 1.12-1.55-1.38 SGPP2.76 1.3-1.77-1.71 C12orf75.7-1.54 1-1.54 PTPN3.17-1.77-1.6-1.88 RDH1.35-1.2-1.75-1.79 DUSP2.4 1.92-1.72 1.12 LOC1537544.42 1.13-1.56-1.38 RAB8B.3 1.2-1.81-1.77 CCL3L3.72 2.17-1.81 1.2 ADO.69 1.7-1.52-1.42 SH2B3.2 1.41-2.1-1.49 CHRNA6.7-1.79 1.6-1.69 TIGIT.35 1.19-1.6-1.34 CXorf21.16-1.42-1.49-2.1 MFSD2A.11 1.75-1.69 1.4 STAP1.24-1.76-1.7-1.88 P2RY1. -1.32-1.56-2.6 XAF1.7-2.1 1.11-1.81 STAT5A.1 1.2-1.61-1.34 MPZL3.7 1.19-1.97-1.65 IGKV3D-15.11-1.77 1.2-1.74 RILPL2. 1.35-1.7-1.26 CD38.49-1.25-1.62-2.2 GSTA4.22 1.74-1.74 1 LOC1537498.99 1.23-1.63-1.33 FSCN1.2-1.24-1.63-2.3 CCDC141.9-1.91 1.5-1.83 LOC15373315.3 1.8-1.54-1.43 TIAM2. 1.48-2.19-1.48 IRF8.8 3.2-2.58 1.17 PHLDA1.71-1.3-1.46-1.51 CD7.1 1.29-2.7-1.61 IFNG.21 2.9-2.89 1.1 NELL2. -1.38-1.25-1.73 ACTN1.21 1.6-1.89-1.79 MDFIC.14 1.45-1.52-1.5 EBI3.5 1.48-1.78-1.2 SESN3.9-1.3-1.82-1.88 LOC1129973.2-1.46-1.5-1.53 FAM15A.1-1.69-1.11-1.87 CD83.25 1.59-2.27-1.43 B3GNT5.42 1.42-1.53-1.8 EGR2.12 1.37-1.73-1.26 BCL2L1.1 1.4-2.17-1.55 ARHGEF2.49 1.33-1.5-1.13 NINJ1.14 1.24-1.66-1.34 TRAF1. 1.29-2.1-1.63 LOC15371864.1 1.34-1.51-1.13 NRIP1.25-1.18-1.38-1.62 DENND5A.13 1.1-1.95-1.78 IL9R. -1.52-1.5-1.59 PPFIBP1. -1.57-1.17-1.84 PRRT3.3 1.2-1.91-1.88 ADAP1.14 1.38-1.53-1.11 LINC158.59 1.94-1.99-1.2 MIR146A.2 1.12-2.1-1.79 PIKFYVE.58-1.38-1.11-1.53 GPR174.26-1.24-1.35-1.67 LOC112934.36 1.4-1.94-1.86 EED.18 1.62-1.65-1.1 LRIG1.2-1.3-1.5-1.54 PELO.46-1.4-1.59-2.22 RAB11FIP1.69 1.33-1.52-1.14 IGFLR1. 1.18-1.64-1.4 ZNF74.5 1.75-2.5-1.43 ACSL6.1 1.66-1.67-1 FGFR1.1-1.41-1.26-1.78 AICDA.45 1.8-2.5-1.9 DOK2.4 1.32-1.52-1.15 NFKB2.4 1.2-1.66-1.38 CCL4L2.14 2.67-2.9-1.9 ADRBK2.22-1.41-1.11-1.56 CYSLTR1.46-1.6-1.17-1.88 RGS16.28 1.47-2.37-1.62 IQCF2.27-1.36-1.13-1.54 STARD4.34 1.9-1.59-1.46 CCR8.8-1.54-1.57-2.43 EPB41L4B.37-1.3-1.16-1.51 ADGRE5.4 1.4-1.56-1.5 MIR2C.52-1.28-1.78-2.27 TP53I11.46-1.37-1.13-1.54 AGK.1 1.18-1.66-1.41 CRTAM.4 1.31-2.33-1.78 C16orf45.71 1.63-1.66-1.2 NFKBIA.4 1.41-1.8-1.27 CD16.7 2.83-3.4-1.8 GCNT2.39-1.33-1.15-1.53 CAV1.1-1.9-1.47-1.61 LAYN.1-1.23-1.86-2.29 BMF.3 1.34-1.55-1.15 GUCY1B3.14-1.37-1.3-1.78 BACH2.1 1.39-2.43-1.75 SIAH2.3 1.33-1.54-1.16 PLPP1.1 1.2-1.56-1.53 PTGIR.2 1.29-2.36-1.82 ZBTB32.46 1.69-1.71-1.1 TNF.56 1.67-1.94-1.16 TAGAP. 1.95-2.78-1.43 CD52.21-1.26-1.2-1.52 ZG16B.1-1.37-1.31-1.79 TBC1D4.1 2.12-2.89-1.37 LINC853.39 1.26-1.52-1.21 TRGV1.7-1.58-1.21-1.9 SPRY1.33 1.59-2.66-1.67 LOC44867.83-1.35-1.16-1.57 SERPINE2.58 1.57-1.9-1.21 MYO1E.9 1.45-2.58-1.78 TTN.47-1.5-1.1-1.65 TRIP1.1 1.41-1.82-1.29 REL.32 1.42-2.64-1.86 COL6A2.53-1.29-1.2-1.55 AHI1.63-1.31-1.35-1.77 CD8.91 2.19-3.9-1.41 VIM.13-1.51-1.1-1.66 SGK223.8 1.34-1.79-1.34 PHEX.15 3.33-3.53-1.6 GBP4.22-1.74-1.1-1.75 CD19.1 1.62-1.95-1.2 LOC1537177.6 1.41-2.73-1.94 IFNAR2.1-1.45-1.13-1.64 HVCN1.6 1.35-1.82-1.34 NHS.1-1.19-2.16-2.56 TSPAN13.81 1.41-1.62-1.15 SHC4.19 1.81-2.4-1.13 CCL4.8 2.69-3.49-1.29
vs 28z vs BBz KIAA1324.4-1.3-2.38-2.45 VCAM1.58-2.42-1.41-3.42 ABTB2.17 1.53-2.98-1.94 IL23A.12 2.34-3.45-1.48 XCL1.31 1.38-2.88-2.9 DHRS3. 1.51-3.1-1.99 PDE7B.1-1.88-1.74-3.27 IGF1.1 1.64-3.18-1.94 IL13.34 1.11-2.71-2.43 TNFRSF9.12 2.14-3.59-1.68 SDC4.2 1.93-3.53-1.83 IL1A.12 2.39-3.8-1.59 NR4A2.4 2.14-3.76-1.75 DUSP1.1 1.2-3.2-2.94 IER3.1 1.69-3.78-2.23 NR4A3.7 2.47-4.58-1.85 HNF1B.3-1.8-3.25-3.51 AFAP1L2.5-1.34-2.96-3.96 CD82.11 1.48-4.2-2.83 CD2. 3.93-5.88-1.49 TNFSF4.9-1.4-4.85-5.3 XCL2. 2.69-1.5-3.9 CSF2.1 3.35-13.94-4.16 CCL22. 2.4-12.63-6.18 IL2.3 13.36-18.1-1.35 CCL1.1 2.14-34.25-15.97 IL3.1 13.75-47.44-3.45
72 hours after NALM-6 stimulaiton 1351 genes) vs 28z vs BBz z vs BBz z vs BBz IL18RAP.1 2.7 1.79 4.82 LINC1358.27-1.86 5.15 2.78 HIST1H4B.23 1.1 1.97 2.18 GZMB.4 1.9 1.92 3.66 ITGA6. 1.49 3.9 4.61 LOC154761.1-1.25 2.3 1.83 IL1.95-1.2 2.6 2.56 CATSPERB.12-1.7 4.53 2.66 HYKK.54-1.15 2.19 1.91 NEAT1.1-1.11 2.51 2.28 FGFBP2. -2.47 4.98 2.1 LOC12724275.1-1.33 2.34 1.76 IL12A.5-1.67 2.91 1.74 VSIG1.1-3.14 5.16 1.64 NUCB2.1-1.46 2.44 1.66 MXD1.4-1.9 2.14 1.96 LOC11929355.68-1.7 3.3 3.9 KLRC3. -2.47 2.91 1.18 GAS2L3.9 1.5 1.99 2.9 GIMAP4. -2.23 4.37 1.96 MUC1.41-1.1 2.14 1.95 CCR5.3-1 2.4 2.3 TNFRSF8.17 1.91 2.12 4.5 SEC31B.3 1.16 1.89 2.2 NKG7.4 1.38 1.63 2.26 LINC239. -1.8 3.18 2.96 JUN.32-1.23 2.25 1.83 IFNG.58 1.92 1.33 2.55 CXCR6.1 1.5 2.99 3.14 HELB.4-1.52 2.45 1.61 LPCAT3. -1.2 2.9 1.74 GLUL. 1.9 2.93 3.2 GNLY. -1.94 2.66 1.37 KLRC1.23 1.9 1.8 1.96 CEP112.1-1.52 3.69 2.43 KLRD1. -1.74 2.55 1.47 F2R.1 1.21 1.62 1.96 BCL6.1-2.1 4.8 2.3 RNF213. 1.1 1.91 2.1 PLEKHF1.61-1.7 1.63 1.52 IGKV3OR2-268.4-1.24 3.3 2.66 LGMN.39-1.7 2.7 1.93 CD47.1-1.6 1.62 1.52 TJP2.5-1.23 3.23 2.63 GIMAP2.2-1.64 2.48 1.51 CCNE1.29-1.33 1.77 1.33 LINC861.1-1.92 3.83 1.99 LINC155.8-1.28 2.23 1.75 GLIPR1. -1.47 1.81 1.23 PDLIM1.4 1.8 2.7 2.9 CCDC126.16-1.27 2.22 1.75 PLAC8.5-1.59 1.79 1.12 LY96.3 1.66 2.1 3.49 PLSCR1. -1.7 2.5 1.92 BMPR1A.12 1.26 1.22 1.53 MAP3K5. -1.22 3.7 2.51 SPATS2L.27 1.34 1.7 2.27 PRDM1.64 1.24 1.22 1.51 NUAK2.5-1.47 3.32 2.26 IGKV3D-7.1-1.13 2.1 1.86 SEMA7A.34 1.35 1.15 1.56 ELOVL4.4 1.38 2.29 3.16 LOC11928173.21 1.68 1.48 2.48 ASXL1.9-1.35 1.54 1.14 FBXO32.2-1.79 3.44 1.93 FAM228B.5-1.27 2.2 1.74 ZFP36. -1.6 3.68 2.31 GIMAP7.1-2.51 3.83 1.53 ADD2.1 1.43 1.62 2.31 CD28.4-1.79 3.39 1.9 IL26.53-1.19 2.91 2.45 ABCB1. -1.29 2.21 1.71 SBNO2. 1 2.47 2.47 TRPS1. 1.32 2.31 3.4 PITPNC1. -1.28 2.2 1.72 ABLIM1.1-1.66 2.82 1.7 LOC15373955.11-1.27 2.97 2.34 STRIP2.4 1.6 1.51 2.4 TIAM1.2-1.93 2.7 1.4 IKZF3. -2.9 3.59 1.72 MVP.15-1.18 2.11 1.79 PPP1R3B.11 1.16 1.85 2.15 SLC9A9.2-1.82 3.39 1.87 ARRDC3.1-1.36 2.22 1.64 ITGA3.7-1.4 1.91 1.84 TC2N. -2.17 3.52 1.62 FURIN.1 1.54 1.52 2.34 CD2R1.3-2.7 2.47 1.19 STOM. 1.69 1.89 3.18 GCA.34-1.4 1.97 1.89 TBC1D15.8-1.15 1.81 1.57 PIK3AP1.1-1.3 2.57 2.49 GRAMD1B.12 1 1.89 1.89 METRNL. 1.86 1.17 2.16 TRIB2.1-1.74 3.16 1.82 HIST1H2BC.12 1.5 1.83 1.93 RAPGEF6.6-1.1 1.75 1.58 KCTD6.31 1.13 2.34 2.64 LOC1192792.4-1.52 2.26 1.49 ARHGAP12.5-1.17 1.75 1.5 GIMAP1.9-1.63 3.5 1.87 TXK. -1.26 2.9 1.66 GPSM2.5-1.21 1.77 1.46 ZFYVE28. -1 2.47 2.45 SLC27A2.16 1.28 1.64 2.1 GALNT1.16 1.1 1.6 1.62 TP53INP1.1-6.6 5.91-1.2 ISG2.3-1.6 2.3 1.43 JAK3.86-1.6 1.63 1.54 SELL. -3.33 3.72 1.12 CAPRIN2.2-1.22 2.4 1.67 NPC2.2-1.1 1.66 1.51 ABCA2. -1.4 2.81 2 GUSBP11.4-1.29 2.9 1.62 IMPA2.79 1.9 1.51 1.65 GZMH.1 1.25 2.12 2.65 PDGFRB.4 1.39 1.55 2.16 PGS1.6 1.16 1.44 1.68 SDK2.27-1.22 2.59 2.11 SETBP1.4 1.6 1.43 2.28 CFLAR.1-1.87 1.99 1.7 GAB3. -1.5 2.4 2.29 PTPN13.42 1.2 1.83 1.86 CTSA.4-1.34 1.71 1.28 MIR21.2-1.18 2.53 2.14 LINC88.13-1.14 1.96 1.72 SIGIRR.67-1.34 1.68 1.25 AIM2.7 1.7 2.25 2.41 SH3BP5.4-2.47 2.61 1.6 IQGAP2.1-1.16 1.57 1.35 GIMAP8.1-2.8 3.14 1.51 CSTF2T.5 1.7 1.76 1.9 IL17RA.62-1.25 1.53 1.22 ANKRD2A1.3 1.15 2.16 2.48 CD3A.5 1.61 1.4 2.26 PRPF38B.15-1.27 1.5 1.18 CLU.86 1.12 2.19 2.45 GSDMB. -1.36 2.1 1.54 SOCS3. 1.29 3.15 4.6 KDSR. 1.58 1.79 2.84 LINC158.16-1.66 2.27 1.37 BCL3.4-1.67 2.51 1.5 HSBP1L1.93-1.56 2.8 1.8 CD8B.8-1.15 1.95 1.69 JUNB.5-1.58 2.33 1.47 RARRES3.11-2.5 3.8 1.51 MX2.89-1.25 2.1 1.61 BATF.7-1.7 1.93 1.81 MIR3143.8 1.38 1.9 2.63 JHDM1D.14 1.12 1.7 1.91 DOCK9.2-1.47 1.96 1.33 LOC1537526.1 1.57 1.76 2.76 HIVEP2.2-1.49 2.15 1.45 SUSD3.26-1.16 1.64 1.41 FASLG.3-1.13 2.38 2.11 ACVR1.2 1.12 1.69 1.9 BCR.4-1.14 1.54 1.35 NUGGC.5-1.36 2.57 1.89 CASP8.13-1.53 2.17 1.42 DUSP16.1-1.77 1.84 1.4 STYK1.29 1.41 1.84 2.6 SLAMF6. -1.8 2.31 1.28 DPY19L3.47-1.27 1.6 1.26 C8orf88.82 1.29 1.94 2.5 TMEM65.5 2.36 1.7 2.52 MCTP2.8 1.19 1.3 1.54 PARP9.24 1.1 2.2 2.23 ANKRD2A3.53 1.6 1.73 1.84 NUP98. -1.14 1.5 1.32 LOC389765.97 1.21 2 2.42 KLF1.2 1.5 1.74 1.83 TIPARP.12-1.21 1.52 1.26 FAM184A.24 1.5 2.15 2.25 CMTM8.4 1.11 1.69 1.88 TANC2.1-1.82 9.87 5.41 PPIAL4E.27 1.13 2.5 2.31 GBP2. -1.13 1.89 1.67 SULT1B1.7-1.24 7.82 6.3 LUCAT1.4 1.8 2.8 2.25 CEP7. -1.52 2.14 1.41 KIAA226L.2-1.43 8 5.61 COL6A3.92-1.35 2.47 1.83 LOC1537333.38 1.35 1.51 2.4 LOC1537676. -1.1 4.73 4.29 RFX8.14-2.35 2.99 1.27 PHF11.2-1.8 1.84 1.71 ZNRF1. 1.2 4.46 4.55 INTS1. 1.35 1.79 2.42 LOC1272481. -1.36 2.3 1.49 TRERF1. -1.24 4.94 3.99 RASA3. -1.74 2.66 1.53 LINC884.18 1.11 1.66 1.85 LOC11927613.5-1.4 4.17 4 RRAGD.4-1.38 2.43 1.76 PHOSPHO2.5 1.14 1.64 1.87 RCAN2.4 1.26 3.57 4.48 DUSP1.6-1.67 2.59 1.56 CD7. -1.11 1.84 1.66
vs 28z vs BBz z vs BBz z vs BBz CLEC2B. -1.11 1.84 1.66 ZNF658B.3-1.3 1.82 1.39 UBL3.6-1.87 1.96 1.5 PLGLB1.75-1.25 1.93 1.55 SYCP2.16-1.76 2.4 1.16 LDAH.1-1.15 1.61 1.4 OGFRL1.2 1.2 1.72 1.75 FAM8A1.4-1.37 1.85 1.35 PFKFB3.18-1.21 1.65 1.36 CCDC69.8-1.3 1.76 1.7 GPR16.14 1.32 1.38 1.82 SGMS1.9-1.32 1.71 1.3 SMIM1.11 1.55 1.36 2.1 LOC1289361.33-1 1.6 1.59 FKBP11.3 1.35 1.28 1.72 C17orf62.8 1.6 1.68 1.77 ZNRF2.6-1.36 1.84 1.35 GDPD3.16 1.7 1.45 1.55 GLUD1.15 1.25 1.54 1.91 BLVRB.2 1.11 1.51 1.68 AGO4.3-1.68 1.87 1.12 LOC15729.4 1.9 1.65 1.8 IGFBP7.6-1.13 1.69 1.5 BCAT1.3 1.8 1.7 1.92 SLC2A3.3 1.74 1.26 2.19 ZFP36L2.1-1.85 2.7 1.12 HEATR5A.76 1.16 1.39 1.6 CEP162.2-1.43 2.3 1.42 CLK1.1-1.59 1.95 1.23 TIMD4.16-1.23 1.65 1.34 LOC11929241.12-1.18 1.86 1.58 GRAMD1C.67-1.56 1.94 1.24 ADD3.6-1.31 1.69 1.29 TCAF2. -1.6 1.77 1.67 LOC1128816.3-1.13 1.69 1.49 OMA1.6-1.16 1.6 1.37 WDR59.1-1.5 1.76 1.68 METTL23.6-1.14 1.69 1.49 LDOC1L.77-1.47 1.77 1.2 SETDB2. -1.34 1.97 1.47 MKRN3.5 1.19 1.45 1.73 LOC1192865.77 1.9 1.42 1.55 ZNF683.34-1.31 1.95 1.49 OARD1.64-1.13 1.69 1.49 MTERF2.51-1.12 1.57 1.4 GIMAP6.44-1.92 2.26 1.18 CXorf57.35 1.4 1.55 1.62 OPN3.66 1.4 1.45 1.52 INADL.1-1.17 1.85 1.58 DPY19L1.53 1.2 1.56 1.6 ZDBF2.18 1.26 1.31 1.66 AGTPBP1.1 1.18 1.57 1.85 GPRIN3.13-1 1.58 1.58 ZNF141.14-1.12 1.57 1.4 LAG3.7 1.43 1.41 2.1 ZNF11.1 1.6 1.53 1.63 ZNF35. -1.28 1.67 1.3 FAM46C.19 1.26 1.51 1.9 ATP2B4.1-1.25 1.76 1.4 KLF3.3-1.83 1.91 1.5 PCED1B.13-1.66 2.13 1.28 HENMT1.1-1.22 1.73 1.42 FAM16B1. -1.35 1.7 1.26 PHYKPL. -1.9 1.78 1.63 LOC1192727.32-1.27 1.76 1.39 MAN1A1.12 1.15 1.38 1.58 NARF. -1.5 1.74 1.66 LOC645513.51-1.11 1.66 1.49 PQLC3.14-1.17 1.6 1.36 FLJ32255.23-1.34 1.95 1.45 ZNF84.9-1.31 1.78 1.36 PTPRCAP.8-1.41 1.73 1.23 KLHDC1.41-1.46 2.2 1.38 LOC1272354.84-2.5 2.11 1.3 PRG4.83-1.8 1.9 1.5 RGS9.6-1.59 2.8 1.31 STK17A.8-1.21 1.72 1.42 AQP3.2-1.75 1.87 1.7 VWA5A.17-1.12 1.79 1.6 RAB38.12-1.9 1.63 1.5 SLC16A7.5 1.19 1.34 1.6 GPR135.98-1.7 1.75 1.63 ADGRE1.97-1.35 1.8 1.33 FAM89B.34-1.76 1.88 1.6 LOC15373495.2-1.52 2.4 1.34 JAKMIP2.48-1.34 1.79 1.34 GSAP.12-1.1 1.54 1.4 MT1L.29-1.46 2.1 1.37 UHRF2.64-1.11 1.65 1.48 TLR1.5-1.16 1.58 1.35 RWDD2A.8-1.27 1.89 1.49 TRIM5.2-1.58 1.91 1.21 ZNF44.3-1.3 1.66 1.27 IRAK4.6-1.22 1.85 1.52 ACVR2A.53-1.4 1.82 1.3 DHRS12.4-1.8 1.51 1.41 PAPD5.2-1.14 1.79 1.58 RNFT1.11-1.25 1.73 1.39 GZMM.1 1.1 1.39 1.53 RNF157.9 1.59 1.3 2.7 ST6GALNAC3.52-1.7 1.61 1.5 JMJD1C.16-1.9 1.52 1.4 ENPP2.84-1.65 2.9 1.27 HMGB1.28-1.47 1.85 1.26 TPP1.1-1.45 1.73 1.19 MT1F.22-1.97 2.23 1.13 NOTCH1.1-1.1 1.63 1.48 CHST12.9-1.6 1.79 1.12 FOSB.53-1.39 1.96 1.4 CST7.1 1.37 1.31 1.79 F5.38 1.7 1.41 1.5 PAFAH1B3.14 1.11 1.59 1.76 LOC1996286.16-1.67 1.94 1.16 KDM7A.5 1.18 1.33 1.58 LOC11638.23-1.45 1.98 1.37 DENND6A.8-1.24 1.71 1.39 MYO1F.66-1.34 1.67 1.24 ARIH2.2 1.35 1.42 1.92 INPP4B.52 1.16 1.43 1.67 OASL.84 1.28 1.28 1.63 NEMP2.25-1.9 1.74 1.59 LOC11929947.27-1.12 1.64 1.46 FYB.1-1.55 1.76 1.14 NMRK1.26-1.58 2.4 1.29 ATF7IP2.11-1.78 1.98 1.11 HIST1H2BE.68 1.29 1.27 1.63 CCNL1.26-1.46 1.97 1.36 CYTH4.17-1.51 1.86 1.23 LZTFL1.61-1.8 1.51 1.39 BIN2.1-1.7 2.9 1.23 SLC46A3.3-1.66 1.93 1.16 SENP7.4-1.44 1.71 1.19 BNIP3.4 1.96 1.12 2.19 CD9.2-1.16 1.66 1.43 ITGAX.65-1.79 1.85 1.4 SATB1. 1.12 1.56 1.74 ERMP1.17-1.13 1.64 1.45 ATP8B2.48-1.16 1.55 1.34 HS3ST3B1.2-1.7 2.7 1.22 IFI6.76-1.31 1.75 1.34 TMEM7.26 1.45 1.18 1.71 SNORD111B.22-1.13 1.74 1.55 MSL3.16-1.29 1.74 1.35 EZH1.12-1.55 1.75 1.13 GSDMA.4-1.22 1.81 1.48 ATP6VA1.89-1.25 1.71 1.37 MYLIP.1-1.57 1.76 1.12 GSTM4.59-1.22 1.81 1.48 C5.83-1.42 1.81 1.27 CRIP1.1-1.22 1.58 1.3 ZNF92.46-1.24 1.82 1.47 CREBZF.59-1.14 1.64 1.44 HIST1H2BN.8-1.21 1.58 1.3 THADA. -1.13 1.74 1.54 OLFM2.75-1.33 1.76 1.32 KLHDC7B.56-1.52 1.73 1.14 IFT8. -1.62 2.3 1.25 PDK1.4 1.33 1.32 1.76 MALAT1.15-1.14 1.53 1.34 PICALM.5-1.7 1.69 1.58 FASTKD3.64 1.2 1.52 1.55 MXRA7.52-1.33 1.64 1.23 GNAI3.58 1.7 1.58 1.68 BFSP1.27-1.36 1.77 1.3 TAS2R9.63 1.16 1.33 1.54 IFT88.2 1.3 1.6 1.66 DHX32.2 1.35 1.31 1.76 ABCA5.28-1.15 1.53 1.33 ERI2.28-1.26 1.82 1.44 DTX3L.17 1.1 1.52 1.53 NRSN2.51 1.16 1.32 1.54 CDC25B.1-1.83 2.1 1.15 AAED1.46 1.13 1.43 1.62 POLG2.85-1.12 1.51 1.35 PCCA.16-1.3 1.65 1.6 HIST1H2AL.7 1.34 1.3 1.75 LOC1537187.53-1.39 1.66 1.2 LRRC8C.77-1.66 2.2 1.22 SNTB2.33-1.13 1.62 1.43 ZNF37BP.57-1.38 1.66 1.2 SERPINB1.7-1.24 1.79 1.45 TPT1.3-1.35 1.75 1.3 ZNF91.1-1.41 1.67 1.19 APPL1.4 1.7 1.56 1.67 GOLGA2P5.93-1.18 1.64 1.4 MGAT4A.6-1.32 1.62 1.23 VPS8.24-1.4 1.65 1.58 LRRC39.31-1.14 1.62 1.42 CFL2.82-1.22 1.56 1.28 FAM214A.1-1.35 1.85 1.37 ORMDL3.1-1.31 1.72 1.32 LUC7L.29-1.27 1.59 1.25 CLUAP1.5-1.2 1.63 1.59 LONP2.55-1.73 1.92 1.11 PPP2R5C.1-1.45 1.68 1.16 SSH2.6-1.6 1.66 1.56 LINS1.24-1.13 1.6 1.42 CECR5.55 1.36 1.2 1.63 DTNBP1.8-1.23 1.78 1.44 SIAE.65-1.4 1.54 1.48 MAN2A1.91-1.3 1.6 1.23 MFGE8.1-1.54 1.95 1.26 DKFZP586I142.7-1.68 1.89 1.12 PRMT2.26-1.41 1.66 1.17