SUPPLEMENTARY INFORMATION

Transcript

1 doi: /nature10648 Supplementary Figure 1: Timing of CHIR99021 exposure determines induction of FOXA2/LMX1A midbrain floor plate precursors. a) Immunocytochemical analysis of FOXA2(red)/LMX1A(green) at day 11 of differentiation following LSB/S/F8 treatment alone or in combination with CHIR starting at the various time points indicated. b) Quantification of the percentage of FOXA2+, LMX1A+ and double labeled cells at day 11 of differentiation following differential onset of CHIR exposure as described in (a). Significance levels for individual markers are presented as compared to no CHIR condition: ANOVA; Dunnett test: *** p < 0.001; ** p < 0.01; p < 0.05). Scale bars correspond to 50 μm. 1

2 Supplementary Figure 2: FGF8 exposure does not play a major role in the induction of FOXA2/LMX1A midbrain floor plate precursors. Representative images of FOXA2(red)/LMX1A(green) expression by immunocytochemistry at day 11 of differentiation. Cells were exposed to LSB/ CHIR in the presence or absence of SHH (purmorphamine + SHH C25II) and FGF8. Scale bars correspond to 50 μm. 2

3 Supplementary Figure 3: Exposure to high dose of SHH and/or a smoothened small molecule agonist (purmorphamine) is required for efficient midbrain floor plate induction in the presence of CHIR Representative images of FOXA2(red)/LMX1A(green) immunocytochemistry at day 11 of differentiation. Cells were treated with LSB/F8/CHIR in the presence of various concentrations of SHH (SHH-C25II) and smoothened agonist purmorphamine. Scale bars correspond to 50 μm. 3

4 doi: /nature10648 a LSB Day1 LSB Day0 S/F8 Day1 LSB Day3 S/F8 Day7 S/F8 Day5 S/F8/CHIR Day5 LSB Day5 S/F8/CHIR Day7 LSB Day7 S/F8 Day13 S/F8 Day11 LSB Day11 LSB Day13 S/F8/CHIR Day13 S/F8/CHIR Day11 b Functional annotation p-value (log10) Fold change GO: ~forebrain regionalization 1 1 GO: ~dorsal/ventral neural tube patterning 2 2 IPR005817:Wnt superfamily 3 3 D11 LSB/S/F8/CHIR vs LSB GO: ~negative regulation of neuron differentiation 4 GO: ~neural tube patterning 5 GO: ~Wnt receptor signaling pathway, calcium modulating pathway 6 GO: ~forebrain cell migration 7 SM00097:WNT1 8 GO: ~pallium development 9 GO: ~limbic system development10 GO: ~telencephalon development11 domain:egf-like 4; calcium-binding hsa04340:hedgehog signaling pathway13 13 GO: ~cerebral cortex development14 14 GO: ~neuron fate commitment15 15 D11 LSB/S/F8/CHIR vs LSB/S/F GO: ~regulation of Notch signaling pathway 1 1 IPR015526:Frizzled related 2 2 GO: ~negative regulation of Wnt receptor signaling pathway 3 3 GO: ~developmental induction 4 4 GO: ~inactivation of MAPK activity 5 5 GO: ~negative regulation of neuron differentiation 6 6 GO: ~neuron fate commitment 7 7 GO: ~regulation of Wnt receptor signaling pathway 8 8 GO: ~negative regulation of MAP kinase activity 9 9 GO: ~diencephalon development GO: ~axon guidance 1 1 D25 LSB/S/F8/CHIR vs LSB GO: ~cell fate commitment 2 GO: ~regulation of neuron differentiation 3 GO: ~pattern specification process 4 GO: ~regionalization 5 Homeobox 6 GO: ~regulation of nervous system development 7 GO: ~neuron development 8 GO: ~neuron projection morphogenesis IPR012287:Homeodomain-related D25 LSB/S/F8/CHIR vs LSB/S/F GO: ~neuron fate commitment 1 1 domain:egf-like domain:egf-like GO: ~endocrine system development 4 4 GO: ~neuron migration 5 5 domain:egf-like GO: ~regulation of neurogenesis 7 7 domain:egf-like Homeobox 9 9 GO: ~forebrain development GO: ~transcription factor activity c Oculomotor Nuclear Complex Red Nucleus Substantia Nigra Ventral Tegmental Area TFF3 EBF1 EBF3 TTR 4

5 Supplementary Figure 4: Analysis of genes differentially expressed in LSB/S/F8/CHIR treated versus LSB and LSB/S/F8 treated cultures at days 11 and 25 of differentiation. a) Hierarchical clustering of the global gene expression data obtained from all three conditions at days 0, 1, 3, 5, 7, 11 and 13 (samples were assessed in triplicates for each condition and each day). b) Gene enrichment analysis according to GO classes using DAVID; Comparisons at day 11 reveal enrichment for both SHH and WNT signaling in LSB/S/F8/CHIR condition in agreement with CHIR99021 mediated activation of canonical WNT signaling. Alternative cell fates such as forebrain development and diencephalon as well as homeobox are other GO terms highly enriched at days 11 and 25. c) Online validation using the Allen gene human expression data base ( for candidate markers enriched in midbrain DA neuron condition (LSB/S/F8/CHIR). TTF3, EBF1, EBF3 and TTR are expressed based on available human brain region specific microarray data. TTR, a classic transcriptional target of FOXA2 is only weakly expressed in adult substantia nigra, suggesting that its main role may be during development or that SHH treatment may cause artificially high TTR expression levels during in vitro differentiation. 5

6 Supplementary Figure 5: Validation of floor plate induction and midbrain DA neuron differentiation protocol in representative independent hesc and hipsc lines. Data from the hesc line H1 and the hipsc lines 2C6 27 (from a sporadic PD patient) and SeV6 (Sendai-based, integration-free) are presented. The FP based protocol described in Figure 1d and Supplementary Figure 6, was used followed by analysis of FOXA2 (red) expression at day 11 and TH(green)/FOXA2(red) at day 25 of differentiation. 6

7 Supplementary Figure 6: Schematic summary of the differentiation conditions used for floor plate-derived and rosette-derived DA neuron cultures. Both protocols used dual-smad inhibition 13 to accelerate neural fate acquisition. LDN was used for BMP inhibition in the FP protocol while the traditional noggin induction was used for rosette cultures. The abbreviations are: LDN: LDN193189, SB: SB431542, SHH (purmorphamine + SHH C25II), BAGCT 2 : BDNF + ascorbic acid + GDNF + dbcamp + TGFβ3. SHH/FGF8 in the rosette protocol used SHH C25I alone in the absence of Purmorphamine following the initial recommendation for patterning of rosette-derived DA neuron cultures. Note: Purmorphamine treatment at rosette stage shows toxicity at concentration suitable for patterning FP cells. BASF: BDNF + ascorbic acid + SHH/FGF8. 7

8 Supplementary Figure 7: In vitro maturation of floor plate-derived DA neuron cultures. a) Immunocytochemical analysis of FP-derived TH neurons (green) for DAT expression (red; day 80). b) Extensive Tuj1+ (red) fiber tracts were observed by day 60 of differentiation extending over distances of > 2 mm. c) At day 80 of differentiation, co-expression of GIRK2 (red) in TH+ neurons (green). Scale bars correspond to 20 μm in (a) 100 μm in (b) and 20 μm in (c) 8

9 Supplementary Figure 8: Immunochistochemical analysis of short-term (6 weeks) in vivo survival studies in adult intact (unlesioned) mouse striatum (NOD-SCID IL2Rgc null strain). a,b) Analysis of FP derived grafts (day 25 cells; 150 x 10 3 cells/animal). a) Representative image of graft core showing TH+ cells surrounded by TH+ host fibers. An average of 6,200 TH+ cells (n=3) were present in the graft at 6 weeks after transplantation. b) FOXA2 expressing cells (red) were only found in the graft but not in the surrounding host striatum demonstrating graft origin and midbrain identity of the cells. Nearly all TH+ neurons (green) co-expressed FOXA2 (red). However, a considerable proportion of FOXA2+ cells did not co-express TH suggesting that these cells have not yet acquired a mature DA phenotype or represent another FOXA2+ neuronal population negative for DA neuron markers. Scale bars correspond to 50 μm. 9

10 Supplementary Figure 9: Histological analysis of long-term (4.5 months) grafted 6-OHDA lesioned mice (NOD-SCID IL2Rgc null strain) comparing behavior of floor plate- versus rosette-derived grafts. a-d) Analysis of FP-derived grafts. a) Example of one of the largest FPderived grafts. The graft retains a well circumscribed hncam+ (red) cytoarchitecture. b) Robust hncam+ fiber outgrowth was observed at graft periphery. c) Serotonergic (5-HT+) fibers (green) in graft are largely negative for hncam (red) suggesting host origin. d) GABAergic neurons and fibers (green) in graft. e-h) Analysis of rosette-derived grafts: e) Neural overgrowth with compression of host brain tissue. The majority of cells were positive for both NCAM (red) and DCX (green) suggesting neuronal fate. f) NCAM+/DCX+ fibers extended to the nontransplanted contra-lateral side of the brain. g) Within the graft core we observed multiple DCX+ clusters. h) Only very few TH+ cells (green) and fibers were observed at graft periphery and 10

11 nearly all rosette-derived TH+ cells in vivo were negative for FOXA2 (blue). i-k) Immunohistochemical analysis of FP-derived versus rosette-derived grafts at 4.5 months. Representative images are presented for expression of the proliferation marker Ki67 (red) and the neural precursor marker PAX6 (green) (i), expression of FOXA2/DCX (j), FOXG1/hNCAM (j, inset), and the astrocytes marker GFAP (green) (k). Scale bars correspond to 500 μm in (a), 50 μm in (b-d), to 500 μm in (e), 50 μm in (g-h), 50 μm in (i), 40 μm in (j) and 20 μm in (k). 11

12 Supplementary Figure 10: Histological analysis of long-term (5 months) grafted 6-OHDA lesioned SD rats. a) The large majority of hna+ (red) cells expressed Tuj1 (green) indicating neuronal fate identity. b) GFAP+ fibers (green) within the graft did not co-express hna (red) suggesting host origin. c) A few human serotonergic (5-HT+) cell bodies (green) were observed while most serotonergic fibers were host-derived similar to the data in mouse host (Supplementary Figure 9C). d) Additional example of TH+ neurons (green) within graft characterized for expression of hncam (red) to confirm human identity of cells in host brain. Scale bars correspond to 25 μm in all panels. 12

13 Supplementary Figure 11: Histological analysis of floor plate-derived grafts in primate brain. FP-derived DA neurons derived from H9 hescs and from H9-GFP hescs (day 25 cultures; 1.25 x 10 6 cells/tract, 3 tracts/per hemisphere, 6 tracts in total) were grafted into the striatum of adult MPTP lesioned rhesus monkeys (=2). a) Upper left and right panels: High resolution image reconstructions of representative graft sites (1 month after transplantation). Graft cytoarchitecture is illustrated by immunohistochemistry for the human specific marker SC-121 (no cross-reactivity with non-human primate tissues). Lower left panel: SC-121+ fibers extending 13

14 from graft core. Lower right panel: Higher resolution image of SC-121+ cells showing neuronal precursor morphologies. b) Analysis of GFP expression in graft cores further confirmed human identity of the cells complementing the SC-121 data. Note: For both animals, one hemisphere each was grafted with unmarked H9 derived cells while the other hemisphere received cells derived from H9-GFP cells (expression of GFP under control of EF1a promoter 38 ). c) Higher resolution image of the GFP+ graft using DAB detection showing numerous GFP+ cells at graft periphery exhibiting neuroblast morphology. d) Immunohistochemical analyses of areas in the graft core negative for GFP and SC-121. These areas contained large numbers of host microglia based on Iba1 expression (red) as well as few ED1+ macrophages (blue), suggesting persistent inflammation despite cyclosporine immunosuppression. Scale bars correspond to 500 μm in (a, top panel), 50 μm in (a, lower left panel), 20 μm in (a, lower right panel), 2mm in (b), 500 μm in (c, left panel), 50 μm (c, upper right panel), 100 μm (c, lower right panel), 100 μm (d). Supplementary Reference: 38. Placantonakis, D. G. et al. BAC transgenesis in human embryonic stem cells as a novel tool to define the human neural lineage. Stem Cells 27, (2009). 14

15 Supplementary Table 1 Column # Column ID Fold-Change p-value 7304 FOXA E FOXA E SPON E SYT E COL22A E FBLN E TFF E DKK E CAPN E C20ORF E PKDCC E LOC E DDC E LDB E AMOT E DBX E SOX E SILV E SHISA E STK E PLCL E MGST E TSPAN E RHOU E HS E DOCK E MMP E SERPINF E RGS E APCDD E IRX E OSBPL E THBS E

16 14581 LOC E DAB E IRX E LMX1A E STOM E ODZ E GSC E NRCAM E PTCH E SULF E FREM E LITAF E NTNG E NEUROG E NPY E NKX PCDH E TNFRSF E EFEMP E SP E PQLC E SLC20A E SNHG E PKP E MYL E NPTX E NMD E LOC E LOC E SEMA6A EYA NPFFR E CXXC E C4ORF E LOC KIAA1324L E

17 23663 LRIG E FABP5L E RPL13A IL1RAPL E PTN E SNHG E RASL E LOC E SLC16A E LOC E TLE E KCNQ1OT E SPATS2L CMTM SNORD C5ORF E TIGA E BMP LOC E NCRNA E GPM6B E DYM S1PR PLEKHA HS E NTN E IRS LOC RPL E WNT5A E SLC38A E HS E LOC E LOC E ITPR E CHN E

18 32162 TIPARP ADSS C17ORF E CCDC ENC POU3F E LOC MAP1LC3A E GAS E LOC E RSPO E RGS E LOC SLC3A E VCAM E AFAP1L E LOC RNF E EMILIN E ZNF E NUAK NEBL E SALL E VANGL SLC7A E ANXA ZFP36L E LOC E ASNS E WNT7B E MARS WDR SPRY E GABARAPL E COL1A E FILIP

19 24246 METRN E HS E PTGIS E KIAA E BCMO E MXRA E TNFAIP E ZFHX RASL11B DUSP CACHD E CAMKV E FLNC E PRSS E ISYNA E GADD45A E EMID E APLP E SLC7A FZD E EMX2OS E ATF E WNT8B E SST E CALCB E ARHGEF E OTX OLFM E TUBB2A TAGLN E DMRTA E CD200R E LOC E WNT5B E RAB11FIP E LOC E

20 27658 PLCH E FLJ E CBX E ERRFI E TMSB15A E KLHL E SIPA1L E TRIB E LFNG E IER E PTPRZ E BEX TLE HS ALPK E TNFRSF12A E TNRC E ANKRD E LEMD E OR7E156P E MGC E ID E PLP E COL4A E COL11A E CA E NR2E E SPOCK E LIX E TFPI E TRIM E TRH E TOX E C1ORF E DMRT E TMEM E

21 2437 C1ORF E C6ORF E ARX NUAK E WNT2B E PTRF E GCNT E EMP E TBC1D E COL5A E CTNNA E C20ORF E CCL E ZNF385B E HS E LRIG E GLI E STMN E SFRP E ARMCX E CTGF E FAM181A E CLDN E ALDH1A E ALDH1A E VAT1L E CYR E KANK E C9ORF E IGFBP E CRIP E CDH E ANKRD E JAG E ID E ANKRD34B E

22 12850 KCNJ E ETV E HS E ALDH E NRIP E SLC2A E DOCK E NAV E DLL E ZNF E ACTC E ARHGAP E SOX E NR2F E CA E TTYH E EFHD E ANXA E HES E CNTNAP E MAMDC E GAS E SP E SFRP E LHX E EMX E LOC E PAX E DRD E NOS2A E BARHL E NOS E HES E-22 Supplementary Table 1: Gene expression array data of significantly up-regulated and downregulated genes at differentiation day 11 in SHH/FGF8/Chir treated Floor-plate based population over control LSB treated population 22

23 23

24 Supplementary Table 2 Column # Probeset ID Fold-Change p-value SYT E FOXA E COL22A E DKK E GPR E LEF E C20ORF E DDC E HS E LMX1A E INHBE E TPBG E ATF DOCK E STOX E DBX E THBS E C5ORF E SULF E SULF E ZNF E IRX E DDIT E STC E PKDCC E COLEC E TFF E MSX LOC E OTX E LDB E LMO E APCDD E DAB E

25 25760 NEUROG E SPATS2L E AXIN E TNFRSF E DDIT E OTX E PTPN E ARL4A E OSBPL E TSPAN E EFNB GSC E FAM84B E SERPINF E ARHGAP E SP E PSAT E RGS LGI E FZD E MUSTN E MYL E PVRL E ADAMTS E BMP E SPON ODZ E XBP E RBP E IRX E ASNS E C6ORF E IL1RAPL E IRS E FOXJ E RARB E

26 14472 LOC E CITED E STT3B E NPY E EYA PCDH E NTNG E ADSS E FAM107A PTPRM E SYT E FLRT E TMEM E PLCL E EGLN CHAC E NR2F E FOXA E KIAA1324L E CDK HS E LOC E CPNE E DDIT4L SDCBP E FLRT ENPP BMP LPAR E LRIG E ERRFI CDCA SNHG E TRAM KCTD RGL

27 30628 SNHG PPPDE E DPYSL MAP1LC3A E MGC D4S234E E DACH E CPXM E TMEM ACBD E HOOK E COPG2IT E TUBB EPHA E RGS E PRKCH E WBP LY6E E SEPP E KCND E ZNF LOC E NMU MEST E C5ORF E PON E MEG E TCF7L YBX E PTGIS E FHDC E MFNG E FEZ E C20ORF E OVOL E RFTN E

28 19038 LOC E SH3BP SOX LOC E DAAM E TMPRSS E OR7E156P E ITPR E FAM65B E JARID E MOBKL2B E SYT E NDRG E MGST E C18ORF E SOSTDC E CXCL E UCA E IER E FNBP E IFITM E NCRNA E PLAC E NES E STMN GCA E IGSF E LOC E CSRP TACSTD RRAGD EPCAM PTRF E KANK E CD200R E UPK E

29 8911 HS E MMD E ETV E PMP E EFHD E SPRY E CDH E SAT E UPP E C14ORF E HS E LOC E CAV E NYNRIN E LOC MPPED E LAMC E MPPED E LAMC E LOC E ENC E ACVR2A E WDR E RAB E CDH E RGMA E S100A E HAPLN E CHST E ISL E PTCH E TOX E F2RL E TMEM E CCND E ANKRD E

30 5241 DLL E VGF LOC E STC E TFAP2C E MN E PVALB E FLJ E GADD45G E CXXC E GRHL E MBIP E DPPA E TMEM E EPHB E STMN E INSM E KLHL E HS E UBL E LMO E ETV E ARHGEF E SPRY E RAB E CXCR E TRIB E PTPRZ E SPOCK E NKD E MYT E FLJ E SLC2A IGFBP E MAF E PRSS E

31 9923 HS E COL5A E CAMKV E SFRP E KIT E FGFR E HOPX E S1PR E VCAM E DLL E PRODH E LOC E ATP1B E FJX E CDC42EP E IFITM E CDKN1C E CAV E SPRED E SH3GL E HDC E HES E USP E ASCL E SFTA E LIPA E PDPN E FZD E CDH E C6ORF E LIMCH E LEMD E ERP E SCRG E FRZB E KIAA E

32 2380 C20ORF E NKX E SOX E POU3F E GPC E LOC E LOC E DLK E HEY E SMS E SFRP E L1TD E GPR E FZD E NKX E GNG E HS E TTYH E DUSP E FABP E DLX E KCNK E SLC15A E FGF E SOX E HESX E RAX E LOC E LOC E LHX E SIX E IFITM E NKX E IGFBP E HES E SIX E

33 Supplementary Table 2: Gene expression array data of significantly up-regulated and downregulated genes at differentiation day 11 in SHH/FGF8/Chir treated Floor-plate based population over SHH/FGF8 only treated population. 33

34 Supplementary Table 3 Column # Column ID Fold-Change p-value 1209 ASCL E CHGA E STMN E TH E STMN E GAP E D4S234E E HES E COL3A E HS E EBF E SCG E INSM E POSTN E PTPRO E SLC17A E SYT E SLC18A E EBF E DCX E C1QL E INA E NR4A E RTN E HS E ADCYAP E BEX E ZCCHC E FLJ E STMN E GNG E NEFM E ID E

35 29017 REEP E SNAP E MYT E UBE2J E DLL E RGS E PPP2R2B E PCSK1N E TFF E PCSK E CRIP E IL13RA E TMEM E SNAP E KIF5C E LOC E TMEM E CDK5R E EEF1A E TNRC E DNER E FAT E ETS E NBEA E KCNJ E CHGB E MXD E CGNL E ONECUT E C6ORF E ID E NHLH E C14ORF E HS E GRM E GADD45G E

36 4298 CNTNAP E AUTS E LRRC4C E SRRM E CHN E PITX E KLC E PLEKHA E MYLIP E DDC E TUBB E MIAT E PEG E TMEFF E SCG E SYP E HS E XKR E TUBB2A E INSM E KIF1A E NAPB E SYT E ELAVL E LMO E CLASP E C7ORF E UCHL E RUNDC3A E MAP1LC3A E ENC RAB40B E MLLT E FRMD4A E KIAA E KLHL E

37 7988 GNB E TAGLN E HS CRMP E LOC E ST6GALNAC E PAFAH1B E TMEM170B E PKIA E BSN EVL E ZMIZ E LOC E TERF2IP E DACH E DNAJC E GPM6A E LOC E HIST1H4K E PRICKLE E SRGAP E LHFP CADPS E CALCA E BMPR E NDRG C18ORF E CTNNA E BEX E TPH E TCEAL E CBLN E RUFY E CELSR E CADM E LOC E

38 13713 LEMD E GPR E TBC1D E GOLSYN E TSHZ E ELAVL IRX E ATCAY E SYT E RIMBP E RAB6B PHF21B E MAPT E HS E HIST2H2BE MAP1B KIAA E ZBTB E RPRM YPEL E LANCL E APLP E SCN3B E RIMS E PDZD ATP6V1G SYT E TMEM106A GATS E AGAP E MARCH E DLL E EVI5L E CALM E SPRY E TTR E

39 33765 WDR E ACPL E DKK E LRRN E TMSL E SCG E TCF E CCDC E DUSP E LOC E NRSN NANOS E DPYSL EXOC E MAPRE E HS E BEX E FAM36A E LBH E HS E WSB E HS E LOC PDCD FZD E KIAA E CNTN E TMEM200A E FAM89B E LOC E ATP9A E GPR137C LOC E CYTH E LOC E C1ORF E

40 21820 LOC E LOC PAPSS TIMP E PIP5K2B HSBP E TP53INP E REM ZNF E CENTA E LY6H RSBN E BRP44L E C3ORF E JUN E LOC KIDINS TUBB4Q ALPP E LOC E LOC FLJ LOC E FLJ E LOC E HS STOX E LOC TRIM E SERTAD NOL PIK3R E PAK E PMP E PINK E CAMSAP E

41 9323 HS E LOC F E C18ORF E ANKRD E DMRTA E FAM117B E AMY1A GABRR E BAZ2B E GLRA E HS E GDAP E SPHK E ST E LOC E LOC SKP E C3ORF PHYHIPL E PDE4D E ANK E ARL E MNX E FLJ E SERINC E ZSWIM E LOC LOC E LOC E GNAO LOC APPBP E NAP1L E LOC E MYCBP

42 27083 PAM LOC SARM E GABRB E SIAH E PITX LOC FAM46A E NT5C ARID5B AFF E RGS E HS E CDH HS E ZNF E HIST1H4J E HS E ALG E VASH RALGPS EFNB BSCL E RHBDL E CDH MAP1LC3B E TMEM E TUBA3E E AGPAT E PLXNA ZMPSTE ZNF RAB9B E CCBE E ELAVL E KIAA E

43 9496 HS ATP6V1B FAM181B E MKL PNMA LRRC37B E FGD E BSDC POTEF C7ORF E SEC11C E SSX2IP E RNF E SLC12A GRK HEPACAM E BEXL E CMIP SHANK LRP1B E FBXL E NLRP LOC C20ORF E TMX E LOC E NCOA E C5ORF E CASD E PRKAR1A EVI E FJX E SORBS E CHD E ZNF CYCS E

44 11686 HS ZADH E LOC E PFN E PARP E DYNC1I GABARAPL AASDHPPT E C12ORF SVOP E ATXN E LOC MIB E TCEAL HSD17B E PDZRN E FAM13B E ATP6V0C E JAKMIP E ZMYND STMN RHBDD E NCOA SLAIN E C17ORF LOC E SND LOC E EXOSC LOC E PPIH E LOC E TYK P4HA E CRTAP E RHBDF E

45 18367 LOC C1ORF E ATP5G MRPL HS E LOC COL2A LOC E LYN LOC TIMM RPA E WDR E NCAPD LOC C14ORF E TYRO E KNTC E LOC ATP6V1B TKT ARHGAP E CYYR E SESN E DDX ITGB3BP E LOC E LOC MAPKAPK HSD17B E TRIM E LOC E STOX RAB E LOC E RPSA

46 17241 LOC E HES RPLP E FGFR E DBX RPS RIPK LOC E PLOD HNRNPA E FHL E LOC E TMEM45A LOC E BAIAP2L E CLEC2D FLJ LOC E LOC E MRPS E LOC SNRPA E CDCA NOB E CDCA WDYHV LOC E LOC LOC E RPL E LOC LOC E RPL E LOC DYM E FANCG E

47 26223 NPY E LOC PHGDH LSM LOC E LOC RPL14L E LOC E EBPL E C4ORF CPNE E LOC LOC LOC E LOC E RPL13L E PDK E SYPL LOC E RPL E LOC E C6ORF LOC RAI LOC LOC ECT E RPS LOC LOC MAD2L LOC STK E SNORD E ATIC TTK E

48 23352 LOC HJURP E LOC E RPP E LOC E LOC RPS TMEM E GALK E PCDH CNTNAP E LOC E EIF3H HNRPC SLC27A E EEF1D SLC44A CENPN RPL LOC PPAT MTP E CCNA WNT5A E PPP1R3C E BMP E LOC E ADA E LGMN E HSP90AB E EIF4B E LEPREL LOC E RPS E CDC PABPC

49 33241 TYMS SHC LOC E EIF3M LOC RPL10A MCM LOC DDIT4L AHCY LOC E EEF1G EFHD E TNFRSF10B E NME1-NME E LOC E HNRPA1P E LOC E PTPN LOC E DARS E C11ORF E LOC E LOC E LOC E CHEK E AKR1A LOC E STOM E LOC RPL SEC11A LOC E RPL13A CDKN SLC20A E

50 13757 LGTN E TNFRSF E SP E MAD2L IMPA E LOC RPA LOC E RPUSD E LOC E LOC E NME HNRPA1L E BCAR LOC E LOC CA TET E HSD17B CPS E MTA E EFEMP E PQLC SHMT LOC E ISG20L E RPS LOC E LOC E MAP4K E GAPDH C1ORF E C18ORF BUB E EIF2S E RENBP E

Δείτε περισσότερα