Supplementary table1. List of gene sets with simultaneously altered the enrichment upon SAP18 overexpression and knockdown. Gene sets enriched by SAP18 and reduced by shsap18 SAP18 against LacZ shsap18 against shscrbl Gene set NAME NES FDR q-val NES FDR q-val KEGG_ARACHIDONIC_ACID_METABOLISM 1.376 0.195-1.548 0.127 KEGG_CELL_CYCLE 1.636 0.128-1.480 0.176 KEGG_DNA_REPLICATION 1.529 0.126-1.562 0.143 KEGG_HOMOLOGOUS_RECOMBINATION 1.563 0.142-1.756 0.061 KEGG_HUNTINGTONS_DISEASE 1.413 0.164-1.691 0.083 KEGG_MISMATCH_REPAIR 1.561 0.132-1.452 0.186 KEGG_NUCLEOTIDE_EXCISION_REPAIR 1.425 0.169-1.260 0.412 KEGG_OOCYTE_MEIOSIS 1.932 0.037-1.457 0.191 KEGG_OXIDATIVE_PHOSPHORYLATION 1.530 0.133-1.443 0.185 KEGG_P53_SIGNALING_PATHWAY 1.459 0.159-1.604 0.139 KEGG_PARKINSONS_DISEASE 1.354 0.192-1.840 0.024 KEGG_PURINE_METABOLISM 1.533 0.137-1.532 0.136 KEGG_PYRIMIDINE_METABOLISM 1.573 0.133-1.435 0.179 KEGG_RNA_DEGRADATION 1.521 0.116-1.483 0.189 KEGG_SPLICEOSOME 1.442 0.172-1.587 0.140 Gene sets reduced by SAP18 and enriched by shsap18 SAP18 against LacZ shsap18 against shscrbl Gene set NAME NES FDR q-val NES FDR q-val KEGG_AMYOTROPHIC_LATERAL_SCLEROSIS_ALS -1.227 0.169 0.818 0.144 KEGG_B_CELL_RECEPTOR_SIGNALING_PATHWAY -1.453 0.214 1.158 0.000 KEGG_ENDOMETRIAL_CANCER -1.222 0.210 1.002 0.023 KEGG_FC_EPSILON_RI_SIGNALING_PATHWAY -1.179 0.243 0.791 0.160 KEGG_GNRH_SIGNALING_PATHWAY -1.239 0.145 0.791 0.177 KEGG_LONG_TERM_POTENTIATION -1.184 0.230 1.073 0.000 KEGG_OLFACTORY_TRANSDUCTION -1.500 0.000 1.372 0.000 KEGG_PROTEIN_EXPORT -1.490 0.147 1.062 0.041 KEGG_RENAL_CELL_CARCINOMA -1.273 0.238 0.586 0.042 KEGG_VASCULAR_SMOOTH_MUSCLE_CONTRACTION -1.209 0.220 1.295 0.000 NES: normalized enrichment score (analyzed using KEGG pathway gene sets in the GSEA software); FDR: false discovery rate (only gene sets with FDR<25% are shown).
Supplementary table2. List of genes with simultaneously altered the expression (over 2-fold and P<0.05) upon SAP18 overexpression and knockdown. SAP18 against LacZ shsap18 against shscrbl Gene Symbol Fold change ANOVA p-value Fold Change ANOVA p-value Acat2 2.21 0.00063-4.25 0.033637 Ap3b1 2.32 0.00215-2.7 0.038013 Arl5b 2.16 0.00018-4.99 0.046193 Atp2b1 3.12 0.000127-2.86 0.026344 Atrx 2.8 0.000024-2.49 0.02811 Bfar 3.34 0.000727-2.3 0.049818 Cab39 2.57 0.000366-2.06 0.023788 Cdc14b 4.2 0.000254-2.45 0.002024 Cideb 2.33 0.003048-6.61 0.024463 Clcn4-2 2.98 0.000306-8.02 0.044548 Clip1 2.08 0.000721-2.64 0.025575 Cpne3 3.39 0.000247-2.08 0.032876 Csnk2a1 2.3 0.000365-3.6 0.009736 Cyp2a4 3.02 0.000012-5.57 0.023834 Ddx5 2.36 0.000223-2.31 0.009329 Dhx9 2.26 0.001125-2.11 0.019649 Dnajc3 2.02 0.00239-2.61 0.032006 Efr3a 4.38 0.000398-2.56 0.008339 Eif2d 2.79 0.000017-2.04 0.036625 Enpp1 2.26 0.000179-2.03 0.039747 Enpp3 2.76 0.000649-2.36 0.04078 Enpp5 3.13 0.000047-2.32 0.03076 Entpd5 2.87 0.000659-2.21 0.032727 Etf1 2.41 0.000002-3.59 0.044613 Fads2 2.13 0.02132-2.12 0.048648 Fam126b 2.84 0.001137-2.38 0.027197 Fth1 2.21 0.00125-3.92 0.028599 Gcc2 4.3 0.000248-2.46 0.032073 Gclc 3.52 0.004522-2.45 0.007792 Gm10094 3.96 0.000012-3.76 0.000312 Golga4 3.2 0.001965-4.6 0.047409 Hdlbp 4.66 0.000583-2.49 0.021566
Hspa13 2.74 0.000954-2.42 0.0316 Jak1 4.67 0.001115-3.45 0.037914 Jhdm1d 2.6 0.000436-3.95 0.015393 Kif5b 2.84 0.001092-2.03 0.030169 Klf9 3.59 0.000177-4.19 0.024514 Mapkapk5 2.23 0.001317-2.56 0.014868 Mmgt1 2.81 0.00037-2.02 0.012737 Mtmr1 2.12 0.001475-3.01 0.031889 Mttp 3.93 0.000033-2.62 0.020725 Ncl 3.12 0.001387-2.58 0.030047 Odc1 2.4 0.000638-2.09 0.030715 Pcmtd1 3.07 0.004705-13.4 0.033587 Pggt1b 2.37 0.000175-2.2 0.003305 Phf17 2.18 0.001007-2.53 0.029462 Ppargc1a 4.08 0.000413-3.18 0.032454 Ppp2r5c 3.2 0.001273-2.09 0.025684 Ptplad1 2.34 0.004689-2.21 0.016007 Rbm25 2.2 0.00419-2.23 0.040231 Rnf8 2.04 0.00073-2.66 0.044214 Rora 3.68 0.001167-2.48 0.011351 Rps24 2.08 0.004845-2.49 0.026564 Scaf11 2.03 0.001606-2.43 0.042681 Secisbp2l 4.16 0.000004-2.35 0.039062 Sfxn2 4 0.000025-3.79 0.007153 Slc25a30 4.25 0.007716-2.9 0.034492 Stt3b 2.73 0.000145-3.38 0.008534 Tbl1x 2.69 0.000708-3.11 0.042436 Tmem176a 2.48 0.001133-3.05 0.021966 Tnpo3 2.12 0.002357-3.42 0.042309 Topors 4.3 0.000218-4.38 0.033318 Usp8 2.05 0.000075-3.33 0.009066 Xpnpep3 3.13 0.001049-4.74 0.042341 Zfp871 2.61 0.006009-3.37 0.039366 Ear12-2.04 0.000123 3.03 0.04746 Fabp7-2.68 0.00063 3.3 0.013331 Gm11428-2.5 0.000065 2.41 0.045565
Lyz2-2.24 0.000444 2.02 0.024729 Mbl1-2.25 0.000055 2.54 0.047272 Psmc6-2.12 0.009229 2.22 0.034324 Rasgrp2-2.13 0.005033 2.16 0.022482 Saa1-2.03 0.013544 3.95 0.039969 Tardbp -2.08 0.000962 2.88 0.017986
A Rela9ve;expression; level;against;control 4; 3; 2; 1; Sap18;mRNA;levels;in;mice;liver * shsap18; shscrbl; LacZ+Cre; SAP18+Cre; B Sap18 βac9n shsap18 shscrbl LacZ+Cre Sap18+Cre HA6tagged intrinsic Rela9ve;band;intensity 2; 1.5; 1; 0.5; intrinsic;sap18; HA6SAP18; shsap18; shscrbl; LacZ+Cre; SAP18+Cre; Supplementary fig.1. SAP18 expression level in fasting mouse liver administrated by adenovirus vectors. A. Sap18 mrna levels in mouse liver injected with respective adenoviral vector at 8th date post administration. Relative expression levels against internal control 36B4 normalized to the mean value of shscrbl are shown (mean±sd, n=6-8). B. SAP18 protein levels in mouse liver injected with respective adenoviral vector at 8th date post administration are shown by western blotting stained with anti-sap18. The chemiluminescence of the western filter stained with anti-sap18 and anti-βactin were developed using SuperSignal West Femto and Pico Substrate (Life technologies), respectively. Bar graph shows the relative band intensities normalized to the mean value of shscrbl (mean±sd). Abbreviations: shsap18, knockdown vector encoding shrna template (pax-shsap18); shscrbl, control vector encoding non-specific targeting shrna template; LacZ+Cre, control vector encoding LacZ (paxcalnlz2) with paxcalncre; SAP18+Cre, overexpression vector (paxcalnlwtit2-sap18) with paxcalncre. P <0.01 (Student s t-test).
Heart Liver Spleen Lung Pancreas Small;intes9ne Kidney Supplementary fig.2. Representative LacZ expression in a mouse co-injected with paxcalnlz2 and paxcalncre adenoviral vectors at 8th date post administration. The fozen sections of the seven organs were fixed with 0.5% glutarldehyde and stained by X-Gal. The scale bars indicate 100 µm.
(mg/dl) 10 T.Chol (mg/dl) 20 TG shsap18; 8 * * 15 Scrbl; LacZ+Cre; 6 SAP18+Cre; 10 4 2 5 Fed; Fas9ng; Fed; Fas9ng; Supplementary fig.3. Total cholesterol and triglyceride concentrations in plasma at day 4 of adenoviral vector-injected mice. Blood samples were collected from the tail veins of vector injected mice (mean±sd, n=6-8 per group) under conditions of food intake ad libitum (Fed) or after 12 h of starvation (Fasting). Abbreviations: shsap18, knockdown vector encoding shrna template (pax-shsap18); shscrbl, control vector encoding non-specific targeting shrna template; LacZ+Cre, control vector encoding LacZ (paxcalnlz2) with paxcalncre; SAP18+Cre, overexpression vector (paxcalnlwtit2-sap18) with paxcalncre. *P <0.05