Supplemental Figures

Supplementary Fig 1. External validation of GSTO1 with patient survival and relation of GSTO1 expression to tumor stages and grades. A, External validation of GSTO1 with patient survival using Rembrandt and Gravendeel Glioma data sets. B, GSTO1 expression is higher in GBM compared to LGG in TCGA data sets. C, GSTO1 expression increases in KIRC is significantly different between tumor stages and grades.

Supplementary Fig 2. Validation of GSTO1 knockout A, Western blot probing GSTO1 in nt WT and GSTO1 KO cells after 10, 20, 30 passages. B, Reverse transcriptional real time PCR detection of GSTO1 in nt WT and GSTO1 KO single clones. C, Sanger sequence of GSTO1 KO cell lines

Supplementary Fig 3. 3-dimension spheroids of (A) HCT116 nt WT and GSTO1 cells, (B) U-87 MG nt WT and GSTO1 cells

Supplementary Fig 4. Neurospheroid formation of U-87 MG nt WT and GSTO1 KO single clones.

Supplementary Fig 5. and sets up- or down-regulated in U-87 MG, HCT116 and A172 as a result of GSTO1 KO in Bru-seq. A, Genes up- or down- regulated in U-87 MG, HCT116 and A172 as a result of GSTO1 KO. B, Top 10 significant enrichment gene sets modulated by GSTO1 KO in enrichment analysis (GSEA).

Supplementary Fig 6. GSEA plots for highlighted significantly changed gene sets. A, GSEA plots for common significantly changed gene sets overlapped in nascent (Bru- seq) transcriptomic profiles between HCT116 and U-87 MG cells in response to GSTO1 KO. B, GSEA plots for UV response gene sets in nascent (Bru-seq) transcriptomic profiles in U-87 MG GSTO1 KO cells. C, GSEA plots for epithelial mesenchymal transition gene sets in nascent (Bru-seq) transcriptomic profiles in U-87 MG GSTO1 KO cells. D, GSEA plots for interferon production and response gene sets in nascent (Bru- seq) transcriptomic profiles in HCT116 GSTO1 KO cells. E, GSEA plots for tight junction, apical junction and gene sets in nascent (Bru-seq) transcriptomic profiles in response to GSTO1 KO. NES = normalized enrichment score; FDR= false discovery rate

Supplementary Fig 7. DAVID analysis of significantly changed genes in RNA-seq. 24 gene sets were enriched in both HCT116 and U-87 MG cells.

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Supplementary Fig 8. Differential activity of cisplatin, carboplatin and oxaliplatin in HCT116 nt WT and GSTO1 KO cell line

Supplementary Fig 9. Differential activity of select drugs in HCT116 nt WT and GSTO1 KO cell line. First concentrations used for each compound are illustrated and all the compounds were 2-fold diluted. Compounds showing differential activities are highlighed in red.

Supplementary Fig 10. Colony formation of U-87 MG and A172 nt WT and GSTO1 KO cells upon treatment with temozolomide. 500 cell/wells of U-87 MG nt WT, A172 nt WT and GSTO1 KO cells and 2000 cells/well of U-87 MG GSTO1 KO cells were seeded into 96 well plates. Indicated concentration of temozolomide was added the next day. Cells were stained with crystal violet after 7 days of temozolomide treatment.

Supplementary Fig 11. GSTO1 KO cells are more sensitive towards radiation. 500 cell/wells of U-87 MG nt WT and GSTO1 KO cells were seeded into 96 well plates. The next day, cells were radiated with indicated dose of radiation. MTT of the cells were determined after 7 days.

Supplementary Fig12. RNA transcription of all GSTO1 isoforms is downregulated and production is attenuated due to the CRISPR/CAS9 genome editing. A, Isoform expression of GSTO1 in CRISPR treated cell lines in RNA-seq. Bars show mean expression across 3 biological replicates. Error bars represent standard error. B, Abundance ratio of each peptide of GSTO1 probed by protein mass-spectrometry. Triplicate for each sample was used.

Supplementary Fig13. Strong agreement of significantly changed gene sets among GSTO1 KO, siGSTO1 KD and GSTO1 inhibitor C1-27 treatment in HCT116 cells. Significantly changed gene sets (GSEA) overlapped among the three treatment in HCT116 cells. Gene sets with FDR<0.1 were considered significant.

Supplementary Fig14. Knockdown GSTO1 did not change growth rate of HCT116 cells. HCT116 shScramble and shGSTO1 cells were seeded into 96 well plate at 200 cells/well after treatment with doxycycline for 3 days. Doxycycline was added to the cells at day 2 and day 4. Colony formation and MTT assays were determined at day 7.

Supplementary Fig15. HCT116 cells grow slower after GSTO1 knockout or knockdown in medium without glutamine. A. GSTO1 KO cells were seeded into 96 well plate at 200 cells/well in medium with or without glutamine. Cell growth was determined using MTT after 7 days of treatment. B. HCT116 shScramble and shGSTO1 cells were seeded into 96 well plate at 200 cells/well in RPMI without glutamine. Doxycycline was added to the cells at day 2 and day 4. Colony formation and MTT assays were determined at day 7.

Supplemental tables

Table 1. Top 30 up-regulated genes in U-87 MG GSTO1 KO Bru-seq

gene ensemblGene fc log2fc

RNASE4 ENSG00000258818.3 251.8141 7.976215 ALG9 ENSG00000086848.14 20.21923 4.337656 TMEM249 ENSG00000261587.2 13.96941 3.804199 AGRN ENSG00000188157.14 9.409367 3.234098 KRT80 ENSG00000167767.13 9.21308 3.203684 SLC9A3R1 ENSG00000109062.11 8.763223 3.131462 SGSM3 ENSG00000100359.20 6.984794 2.804218 LAMA5 ENSG00000130702.15 6.262307 2.646694 CD9 ENSG00000010278.12 6.0346 2.593258 IFITM10 ENSG00000244242.1 5.924491 2.566691 NUPR1 ENSG00000176046.8 5.795572 2.534951 PDK2 ENSG00000005882.11 5.568363 2.477253 TNFAIP2 ENSG00000185215.8 5.491794 2.457277 RRAD ENSG00000166592.11 5.455361 2.447675 TRIB3 ENSG00000101255.10 5.429262 2.440756 HIST1H2BI ENSG00000278588.1 5.301592 2.406426 MKNK2 ENSG00000099875.14 5.290849 2.403499 TXNDC5 ENSG00000239264.8 5.146783 2.363671 DDIT4 ENSG00000168209.4 5.113496 2.35431 HSPA1B ENSG00000204388.6 4.932361 2.302279 NPFF ENSG00000139574.8 4.90164 2.293265 PDGFA ENSG00000197461.13 4.850207 2.278046 MCAM ENSG00000076706.16 4.837727 2.274329 ADAM8 ENSG00000151651.15 4.771018 2.254297 MXD4 ENSG00000123933.16 4.767134 2.253122 LIME1 ENSG00000203896.9 4.595306 2.200161 MIEF2 ENSG00000177427.12 4.525257 2.178 CYBA ENSG00000051523.10 4.470379 2.160397 SAMD11 ENSG00000187634.11 4.443104 2.151568 CHAC1 ENSG00000128965.11 4.429486 2.147139

Table 2. Top 30 down-regulated genes in U-87 MG GSTO1 KO Bru-seq

gene ensemblGene fc log2fc

IPO4 ENSG00000196497.16 -88.72 -6.471 CHMP4A ENSG00000254505.9 -17.68 -4.144 ADAMTS18 ENSG00000140873.15 -15.268 -3.932 C8orf48 ENSG00000164743.4 -11.636 -3.541 RARB ENSG00000077092.18 -10.189 -3.349 NR0B1 ENSG00000169297.7 -7.8689 -2.976 THBS1 ENSG00000137801.10 -6.842 -2.774 NID2 ENSG00000087303.17 -6.7075 -2.746 VGLL3 ENSG00000206538.8 -6.5068 -2.702 SP140 ENSG00000079263.18 -6.2005 -2.632 SIGLEC15 ENSG00000197046.11 -5.9444 -2.572 LMO7 ENSG00000136153.19 -5.7832 -2.532 EFNB2 ENSG00000125266.6 -5.5506 -2.473 KCTD4 ENSG00000180332.6 -5.4216 -2.439 RNF133 ENSG00000188050.2 -4.9113 -2.296 CD274 ENSG00000120217.13 -4.8502 -2.278 U2AF1L4 ENSG00000161265.14 -4.7602 -2.251 CLN5 ENSG00000102805.14 -4.6937 -2.231 BCL11B ENSG00000127152.17 -4.643 -2.215 FAP ENSG00000078098.13 -4.2768 -2.097 MFSD14A ENSG00000156875.13 -4.2656 -2.093 UQCR10 ENSG00000184076.13 -4.1306 -2.046 GLIPR1 ENSG00000139278.9 -4.0732 -2.026 TMEM156 ENSG00000121895.7 -3.9291 -1.974 GNG2 ENSG00000186469.8 -3.9228 -1.972 HIST1H2BB ENSG00000276410.3 -3.8859 -1.958 NOVA1 ENSG00000139910.19 -3.7983 -1.925 GPC6 ENSG00000183098.10 -3.7794 -1.918 RASD2 ENSG00000100302.6 -3.7052 -1.89 PHOSPHO2 ENSG00000144362.11 -3.6268 -1.859

Table 3. Top 30 up-regulated genes in HCT116 GSTO1 KO Bru-seq

gene ensemblGene fc log2fc

ZBTB9 ENSG00000213588.5 7.21285 2.8506 EEF1G ENSG00000254772.9 7.21285 2.8506 LIME1 ENSG00000203896.9 6.63398 2.7299 ZNF177 ENSG00000188629.11 5.7496 2.5235 PAF1 ENSG00000006712.14 5.56774 2.4771 DHRS2 ENSG00000100867.14 5.08547 2.3464 IFI6 ENSG00000126709.14 5.04561 2.335 OASL ENSG00000135114.12 4.8795 2.2867 PKIB ENSG00000135549.14 4.7552 2.2495 TNNT1 ENSG00000105048.16 4.5342 2.1808 IK ENSG00000113141.17 4.38286 2.1319 IFIT1 ENSG00000185745.9 4.36136 2.1248 DAXX ENSG00000204209.11 4.27473 2.0958 OS9 ENSG00000135506.15 4.18806 2.0663 AKAP12 ENSG00000131016.16 4.16955 2.0599 MDP1 ENSG00000213920.8 3.98046 1.9929 PTMA ENSG00000187514.16 3.94896 1.9815 TMEM200A ENSG00000164484.11 3.71201 1.8922 CAMK2D ENSG00000145349.16 3.69695 1.8863 SMARCC2 ENSG00000139613.11 3.59108 1.8444 FAM50A ENSG00000071859.14 3.56696 1.8347 OPTN ENSG00000123240.16 3.55945 1.8317 AHNAK2 ENSG00000185567.6 3.51105 1.8119 PALM3 ENSG00000187867.8 3.43693 1.7811 NAV3 ENSG00000067798.14 3.39743 1.7644 RAB13 ENSG00000143545.8 3.37027 1.7529 FIP1L1 ENSG00000145216.15 3.33534 1.7378 HIRIP3 ENSG00000149929.15 3.28724 1.7169 DDX23 ENSG00000174243.9 3.28395 1.7154 CA11 ENSG00000063180.8 3.25335 1.7019

Table 4. Top 30 down-regulated genes in HCT116 GSTO1 KO Bru-seq

gene ensemblGene fc log2fc

NR5A2 ENSG00000116833.13 -9.1106 -3.188 SYTL2 ENSG00000137501.17 -7.9522 -2.991 F3 ENSG00000117525.13 -5.6306 -2.493 EHF ENSG00000135373.12 -5.0044 -2.323 CCDC89 ENSG00000179071.4 -4.9407 -2.305 RGS2 ENSG00000116741.7 -4.5345 -2.181 ZC2HC1B ENSG00000118491.9 -4.5219 -2.177 CENPS ENSG00000175279.21 -4.4277 -2.147 BDNF ENSG00000176697.18 -4.166 -2.059 BST2 ENSG00000130303.12 -4.1451 -2.051 TMEM249 ENSG00000261587.2 -4.1451 -2.051 SRGAP1 ENSG00000196935.8 -4.1417 -2.05 NECTIN3 ENSG00000177707.10 -3.8507 -1.945 BMP4 ENSG00000125378.15 -3.7877 -1.921 OVCA2 ENSG00000262664.2 -3.3915 -1.762 WLS ENSG00000116729.13 -3.3552 -1.746 PCSK9 ENSG00000169174.10 -3.2892 -1.718 CDRT4 ENSG00000239704.10 -3.2658 -1.707 BBS10 ENSG00000179941.6 -3.204 -1.68 CKLF ENSG00000217555.12 -3.1821 -1.67 HIST1H2AK ENSG00000275221.1 -3.0832 -1.624 ZNF649 ENSG00000198093.10 -3.006 -1.588 ARRDC4 ENSG00000140450.8 -2.9366 -1.554 TMEM86B ENSG00000180089.5 -2.8776 -1.525 TAS2R13 ENSG00000212128.2 -2.8733 -1.523 SDHAF1 ENSG00000205138.3 -2.7347 -1.451 CRIM1 ENSG00000150938.9 -2.7175 -1.442 SYBU ENSG00000147642.16 -2.7007 -1.433 DHFR2 ENSG00000178700.7 -2.6662 -1.415 TLCD1 ENSG00000160606.10 -2.6301 -1.395

Table 5. Top 30 up-regulated genes in A172 GSTO1 KO Bru-seq

gene ensemblGene fc log2fc

KRT19 ENSG00000171345.13 10.7178 3.4219 HIST1H4A ENSG00000278637.1 6.2999 2.6553 PRH1 ENSG00000231887.6 6.29984 2.6553 C19orf33 ENSG00000167644.11 5.39986 2.4329 SNURF ENSG00000273173.5 4.94995 2.3074 HIST1H2AH ENSG00000274997.1 4.49996 2.1699 FDX2 ENSG00000267673.6 4.49988 2.1699 EGFL8 ENSG00000241404.6 3.59991 1.848 HIST1H2BF ENSG00000277224.2 3.47141 1.7955 MC5R ENSG00000176136.6 3.37498 1.7549 HIST2H3D ENSG00000183598.3 3.29996 1.7224 RWDD3 ENSG00000122481.16 2.77377 1.4718 COMMD3 ENSG00000148444.15 2.59999 1.3785 EDN1 ENSG00000078401.6 2.58143 1.3682 TSTD1 ENSG00000215845.10 2.54998 1.3505 DHFR2 ENSG00000178700.7 2.46428 1.3012 ZNF547 ENSG00000152433.14 2.46315 1.3005 SPATA24 ENSG00000170469.10 2.45769 1.2973 RPP25L ENSG00000164967.9 2.42307 1.2768 TIGD2 ENSG00000180346.3 2.42142 1.2759 IGFN1 ENSG00000163395.16 2.36364 1.241 RPS10 ENSG00000124614.13 2.36249 1.2403 NOP10 ENSG00000182117.5 2.358 1.2376 BIVM ENSG00000134897.13 2.31428 1.2106 UCHL3 ENSG00000118939.17 2.31427 1.2106 ANKRD1 ENSG00000148677.6 2.28213 1.1904 EEF1AKMT2 ENSG00000203791.14 2.24999 1.1699 HIST1H1T ENSG00000187475.5 2.1857 1.1281 RGS4 ENSG00000117152.13 2.18318 1.1264 CCDC59 ENSG00000133773.11 2.17143 1.1186

Table 6. Top 30 down-regulated genes in A172 GSTO1 KO Bru-seq

gene ensemblGene fc log2fc

CHMP4A ENSG00000254505.9 -18.12 -4.18 NDST2 ENSG00000166507.17 -6.0398 -2.595 HIST1H2AD ENSG00000196866.2 -6.0398 -2.595 RTL5 ENSG00000242732.4 -6.0101 -2.587 ANG ENSG00000214274.9 -4.4444 -2.152 C11orf98 ENSG00000278615.4 -4.4444 -2.152 NEFL ENSG00000277586.2 -4.3479 -2.12 DAXX ENSG00000204209.11 -4.1799 -2.063 FAM50A ENSG00000071859.14 -4.1207 -2.043 PAF1 ENSG00000006712.14 -4.0617 -2.022 CA11 ENSG00000063180.8 -4.0318 -2.011 CDK5RAP3 ENSG00000108465.14 -3.8889 -1.959 CCDC85B ENSG00000175602.3 -3.7198 -1.895 PTMA ENSG00000187514.16 -3.6879 -1.883 SMARCC2 ENSG00000139613.11 -3.4643 -1.793 OS9 ENSG00000135506.15 -3.3777 -1.756 HOXC5 ENSG00000172789.3 -3.3333 -1.737 ZNF358 ENSG00000198816.6 -3.2889 -1.718 TCEAL3 ENSG00000196507.10 -3.2833 -1.715 CALR ENSG00000179218.13 -3.2226 -1.688 NHEJ1 ENSG00000187736.12 -3.1746 -1.667 FTSJ3 ENSG00000108592.16 -3.1437 -1.652 DCTN1 ENSG00000204843.12 -3.07 -1.618 TM9SF1 ENSG00000100926.14 -3.0556 -1.611 CENPB ENSG00000125817.7 -2.9821 -1.576 FIP1L1 ENSG00000145216.15 -2.9545 -1.563 PRKCSH ENSG00000130175.9 -2.9211 -1.547 BICRA ENSG00000063169.10 -2.9167 -1.544 PIK3R2 ENSG00000105647.16 -2.7778 -1.474 HIRIP3 ENSG00000149929.15 -2.7741 -1.472

Table 7. Commonly up- or down-regulated genes between HCT116 and U-87 MG GSTO1 KO (Bru-seq)

Commonly Up-regulated Commonly Down-regulated genes genes PRKCSH L3HYPDH PTMA TAS2R13 SMARCC2 IPO4 SLC39A7 F3 LIME1 SLC7A11 TRIM21 CYR61 GRIPAP1 HYOU1 PQBP1 SUPT5H OS9 SART1 PPP1R9B CDK5RAP3 UTP14C UBXN1 RNF40 TSPYL2 EHBP1L1 DCTN1 PKM CALR DDX23 SRRT UBTF SF3B2 YJEFN3 CALCOCO1 STX4 TNNT1

Table 8. Up-regulated gene sets enriched in U-87 MG GSTO1 KO Bru-seq (GSEA)

geneset size ES NES pValue fwer fdr

HALLMARK_BILE_ACID_METABOLISM 101 0.39 1.55 0.00635 0.1596 0.042

HALLMARK_DNA_REPAIR 140 0.38 1.58 0.00192 0.1167 0.04

HALLMARK_P53_PATHWAY 187 0.45 1.94 0 0.0006 6E-04

HALLMARK_PEROXISOME 97 0.36 1.44 0.01742 0.4241 0.087

HALLMARK_UNFOLDED_PROTEIN_RESPONSE 108 0.37 1.48 0.01064 0.2911 0.066

HALLMARK_UV_RESPONSE_UP 147 0.44 1.88 0 0.0015 7E-04

GO_ATP_GENERATION_FROM_ADP 33 0.62 2 0.00017 0.1349 0.045

GO_GLUCOSE_CATABOLIC_PROCESS 26 0.65 2.02 0 0.1084 0.054

GO_NADH_METABOLIC_PROCESS 32 0.63 2.06 0 0.059 0.056

GO_NEGATIVE_REGULATION_OF_PROTEIN_BINDING 71 0.51 1.95 0 0.2982 0.082

Table 9. Down-regulated gene sets enriched in U-87 MG GSTO1 KO Bru-seq (GSEA)

geneset size ES NES pValue fwer fdr

HALLMARK_ANDROGEN_RESPONSE 92 -0.35 -1.5 0.006 0.137 0.053

HALLMARK_EPITHELIAL_ 186 -0.41 -1.92 0 0.002 1E-03 MESENCHYMAL_TRANSITION

HALLMARK_KRAS_SIGNALING_UP 165 -0.32 -1.48 0.0023 0.165 0.048

HALLMARK_UV_RESPONSE_DN 141 -0.48 -2.19 0 0 0

Table 10. Top 10 up-regulated gene sets enriched in HCT116 GSTO1 KO Bru-seq (GSEA)

geneset size ES NES pValue fwer fdr

HALLMARK_INTERFERON_ALPHA_RESPONSE 85 0.73 2.79 0 0 0

HALLMARK_INTERFERON_GAMMA_RESPONSE 177 0.59 2.49 0 0 0

HALLMARK_TNFA_SIGNALING_VIA_NFKB 188 0.4 1.73 0 0.0156 0.006

HALLMARK_MITOTIC_SPINDLE 198 0.39 1.68 0 0.0283 0.009

KEGG_TIGHT_JUNCTION 116 0.47 1.89 0 0.0311 0.015

HALLMARK_APICAL_JUNCTION 177 0.37 1.6 0 0.0643 0.016

KEGG_ANTIGEN_PROCESSING_AND_ 58 0.54 1.95 0 0.0117 0.016 PRESENTATION

GO_INTERFERON_GAMMA_MEDIATED_ 55 0.67 2.4 0 0 0 SIGNALING_PATHWAY

GO_RESPONSE_TO_TYPE_I_INTERFERON 53 0.66 2.32 0 0.0002 1E-04

GO_NEGATIVE_REGULATION_OF_TYPE_I_ 36 0.69 2.23 0 0.0009 4E-04 INTERFERON_PRODUCTION

Table 11. Down-regulated gene sets enriched in HCT116 GSTO1 KO Bru-seq (GSEA)

NE geneset size ES pValue fwer fdr S

HALLMARK_MYC_TARGETS_V2 58 -0.45 -1.6 0.00635 0.0709 0.057 GO_ISOPRENOID_BIOSYNTHETIC_ 23 -0.68 -2 0.0002 0.1125 0.087 PROCESS KEGG_GLYCOSYLPHOSPHATIDYLINOSITOL - 24 -0.59 0.00357 0.1588 0.054 _GPI_ANCHOR_BIOSYNTHESIS 1.75 KEGG_N_GLYCAN_BIOSYNTHESIS 46 -0.5 -1.7 0.00449 0.252 0.06

Table 12. Up-regulated gene sets enriched in A172 GSTO1 KO Bru-seq (GSEA)

geneset size ES NES pValue fwer fdr

KEGG_DRUG_METABOLISM_ 38 0.62 2.07 0 0.0023 0.002 CYTOCHROME_P450 HALLMARK_INTERFERON_ 88 0.45 1.74 0.00028 0.0109 0.007 ALPHA_RESPONSE HALLMARK_TNFA_SIGNALING_VIA 183 0.41 1.78 0 0.0066 0.008 _NFKB HALLMARK_KRAS_SIGNALING_UP 171 0.38 1.61 0.00031 0.0427 0.019 HALLMARK_INTERFERON_ 178 0.37 1.57 0 0.061 0.02 GAMMA_RESPONSE KEGG_METABOLISM_OF_XENOBIO 39 0.54 1.8 0.00097 0.0844 0.032 TICS_BY_CYTOCHROME_ P450 HALLMARK_HYPOXIA 188 0.33 1.44 0.00286 0.184 0.052 HALLMARK_UV_RESPONSE_DN 141 0.33 1.37 0.01527 0.3319 0.086

Table 13. Top 10 down-regulated gene sets enriched in A172 GSTO1 KO Bru-seq (GSEA)

geneset size ES NES pValue fwer fdr

HALLMARK_MITOTIC_SPINDLE 198 -0.53 -2.13 0 0 0

HALLMARK_E2F_TARGETS 196 -0.5 -2.03 0 0.0002 8E-05

HALLMARK_G2M_CHECKPOINT 194 -0.49 -1.99 0 0.0004 1E-04

HALLMARK_MYC_TARGETS_V1 196 -0.43 -1.76 0 0.0093 0.002

KEGG_SPLICEOSOME 122 -0.53 -2.01 0 0.0019 0.002

HALLMARK_TGF_BETA_SIGNALING 54 -0.51 -1.71 0.00049 0.017 0.003

GO_RETROGRADE_VESICLE_MEDIATE 73 -0.61 -2.14 0 0.0028 0.003 D_TRANSPORT_GOLGI_TO_ER

GO_SISTER_CHROMATID_SEGREGATI 162 -0.52 -2.05 0 0.0228 0.006 ON

GO_POSITIVE_REGULATION_OF_CHR 79 -0.58 -2.06 0 0.0183 0.007 OMATIN_MODIFICATION

GO_SISTER_CHROMATID_COHESION 99 -0.56 -2.08 0 0.0122 0.007

Table 14. Top 30 up-regulated genes in U-87 MG GSTO1 KO RNA-seq

gene FC log2FC shrunkLFC pValue qValue NELL2 1288.698 10.3317 10.07436 2.66228E-18 3.11723E-17 ZNF883 180.7958 7.498217 7.357162 4.7407E-24 7.54797E-23 PARM1 107.478 6.747898 6.737988 4.9117E-287 1.0837E-283 CALCB 93.00849 6.539291 6.509414 6.11599E-94 7.37932E-92 CYP27A1 65.75752 6.039084 5.8796 1.78129E-18 2.11131E-17 DLX3 58.26247 5.864495 5.82446 1.25053E-63 8.21841E-62 DUSP27 40.04054 5.323389 5.267838 1.75626E-42 5.97439E-41 S1PR1 34.73442 5.118294 5.072912 1.27226E-49 5.53488E-48 JUP 32.98655 5.043806 4.816409 1.99114E-11 1.42631E-10 TMEM92 29.84013 4.899182 4.779922 3.04761E-19 3.78051E-18 SEMA6D 28.32391 4.823949 4.802954 8.97679E-95 1.1003E-92 SELENOP 25.04435 4.646413 4.592613 2.11432E-38 6.23159E-37 NPDC1 24.53137 4.616556 4.609 9.3643E-258 1.2052E-254 NTS 23.61083 4.561377 4.170089 6.98251E-07 3.13118E-06 BICDL1 21.96395 4.457066 4.371356 7.64617E-24 1.19643E-22 ABCA3 21.82817 4.448119 4.421447 1.42173E-72 1.12026E-70 ANGPT2 19.80142 4.307532 4.28275 2.92485E-73 2.42857E-71 HDHD3 18.1204 4.179543 4.083262 2.8791E-20 3.76183E-19 CXADR 16.21602 4.019348 4.008834 5.5765E-161 1.9573E-158 ENPP5 15.86001 3.987322 3.981764 4.6166E-300 1.1883E-296 RPH3AL 12.42049 3.63465 3.57831 8.58576E-29 1.70654E-27 ACTRT3 12.36598 3.628305 3.514315 2.39839E-15 2.28788E-14 PDE1A 12.2149 3.610571 3.503378 3.84547E-16 3.89184E-15 HPGD 12.05107 3.59109 3.562573 1.86169E-55 9.7135E-54 CYP17A1 11.57647 3.533123 3.378546 1.28732E-11 9.37803E-11 ATP1A2 11.2813 3.495861 3.468474 5.05248E-56 2.67228E-54 RCAN2 10.93961 3.451489 3.437383 4.2036E-104 6.2423E-102 DNAH11 10.43394 3.383211 3.371617 1.2182E-123 2.5425E-121 APOL1 9.979545 3.318974 3.305383 1.1494E-103 1.6906E-101 COL3A1 9.97395 3.318165 3.289574 1.62658E-50 7.19797E-49

Table 15. Top 30 down-regulated genes in U-87 MG GSTO1 KO RNA-seq

gene FC log2FC shrunkLFC pValue qValue ZDHHC4 -1734.2 -10.76 -11.96 1.255E-19 1.589E-18 RAB4A -1248.2 -10.29 -11.4206 5.399E-18 6.24E-17 IGFBP1 -845.76 -9.724 -10.7755 4.086E-16 4.124E-15 FOXRED2 -560.55 -9.131 -9.05394 1E-52 4.783E-51 ZIC2 -443.78 -8.794 -9.68179 3.877E-13 3.166E-12 EXO5 -292.41 -8.192 -7.84098 9.71E-12 7.155E-11 GAS6 -196.37 -7.617 -7.55757 1.998E-65 1.353E-63 ELFN2 -177.23 -7.469 -7.41789 1.448E-62 9.203E-61 KRBOX1 -148.01 -7.21 -7.74245 1.543E-08 8.339E-08 PPP1R1C -57.539 -5.846 -5.22191 8.337E-06 3.286E-05 NNT -52.54 -5.715 -5.70141 1.27E-175 5.03E-173 IGFL3 -50.514 -5.659 -4.96284 1.818E-05 6.819E-05 CHST1 -42.753 -5.418 -5.32605 6.158E-27 1.118E-25 ICAM5 -28.526 -4.834 -4.79313 1.292E-52 6.141E-51 GATA2 -26.756 -4.742 -4.64525 7.112E-23 1.061E-21 FAHD2B -21.341 -4.416 -0.26904 0.0112826 0.0257764 TSPAN15 -19.338 -4.273 -4.22411 5.13E-39 1.538E-37 PLEKHG3 -17.66 -4.142 -4.10012 3.798E-44 1.374E-42 APOBEC3G -15.334 -3.939 -3.8775 2.16E-29 4.412E-28 ME3 -14.755 -3.883 -3.70786 1.991E-11 1.426E-10 CABYR -12.689 -3.666 -3.44758 4.75E-09 2.711E-08 DIAPH2 -12.191 -3.608 -3.57267 4.848E-46 1.872E-44 ACAT2 -11.737 -3.553 -3.49788 1.295E-29 2.681E-28 PADI2 -11.671 -3.545 -3.53145 8.02E-115 1.51E-112 ADAMTS18 -11.21 -3.487 -3.45078 1.147E-43 4.08E-42 CA3 -10.924 -3.449 -3.32141 3.009E-13 2.473E-12 HES1 -10.703 -3.42 -3.24976 1.964E-10 1.295E-09 KIF21B -10.561 -3.401 -3.38764 6.7E-113 1.22E-110 LMO7 -9.2142 -3.204 -3.19854 1.23E-253 1.36E-250 ZNF283 -9.0204 -3.173 -3.06961 7.164E-15 6.594E-14

Table 16. Top 30 up-regulated genes in HCT116 GSTO1 KO RNA-seq

gene FC log2FC shrunkLFC pValue qValue SYTL5 265.848 8.0545 8.004734 1.373E-73 2.057E-71 CCL5 78.6262 6.2969 6.107882 1.508E-16 2.169E-15 OAS1 28.7656 4.8463 4.825067 2.792E-93 6.017E-91 IFI6 16.0193 4.0017 3.998862 0 0 IFIT1 11.8149 3.5625 3.558154 0 0 LGALS9 11.7662 3.5566 3.51338 8.427E-37 3.911E-35 PARP10 11.7235 3.5513 3.532224 2.093E-83 3.875E-81 PKIB 11.0163 3.4616 3.455887 1.06E-256 2.09E-253 ALDH1L1 10.4767 3.3891 3.338208 2.565E-30 8.929E-29 KLRK1 10.1995 3.3504 2.334661 0.0039016 0.0105169 IFITM1 10.1505 3.3435 3.326655 2.049E-86 3.883E-84 IFI44 10.0218 3.3251 3.259328 2.513E-23 5.832E-22 EPSTI1 10.0045 3.3226 3.237536 1.519E-18 2.546E-17 GPR15 9.34622 3.2244 3.207155 7.386E-81 1.263E-78 APOBEC3G 9.29796 3.2169 3.008219 2.138E-08 1.366E-07 LEMD1 9.28149 3.2144 3.178729 4.139E-40 2.285E-38 SELENOP 7.70898 2.9465 2.893693 1.366E-25 3.654E-24 IFIT3 7.62316 2.9304 2.912725 1.159E-71 1.635E-69 OASL 7.23152 2.8543 2.841268 3.584E-93 7.62E-91 SERPINF1 7.16047 2.8401 2.747163 7.881E-15 9.873E-14 ITGAX 6.51344 2.7034 2.649306 4.592E-23 1.049E-21 CHAC1 6.43181 2.6852 2.677906 6.76E-155 3.94E-152 TCN1 6.33314 2.6629 2.559687 7.855E-13 8.136E-12 IFIT2 6.30514 2.6565 2.641494 5.377E-76 8.377E-74 SAMD9 5.81917 2.5408 2.534248 9.23E-163 6.31E-160 IFIH1 5.81801 2.5405 2.531432 2.4E-118 7.11E-116 S100A1 5.80545 2.5374 2.423014 2.849E-11 2.489E-10 RAB19 5.77303 2.5293 2.374669 8.725E-09 5.822E-08 DHX58 5.63804 2.4952 2.413854 8.451E-15 1.055E-13 DDX58 5.36858 2.4245 2.418182 2.63E-162 1.73E-159

Table 17. Top 30 down-regulated genes in HCT116 GSTO1 KO RNA-seq

gene FC log2FC shrunkLFC pValue qValue SH3BGRL -2390.5 -11.22 -12.4865 2.454E-21 5.022E-20 WDR72 -921.08 -9.847 -10.9162 1.377E-16 1.984E-15 HMGN5 -868.71 -9.763 -10.8142 3.449E-16 4.817E-15 STXBP6 -781.12 -9.609 -10.6411 8.215E-16 1.115E-14 ZNF607 -603.88 -9.238 -10.2068 1.38E-14 1.687E-13 FANCF -502.78 -8.974 -9.88872 9.402E-14 1.06E-12 ZNF26 -455.56 -8.832 -9.72784 2.271E-13 2.476E-12 ZNF675 -228.11 -7.834 -7.46679 8.854E-11 7.312E-10 ZNF84 -187.6 -7.552 -7.44052 2.594E-30 9.01E-29 ZNF440 -96.002 -6.585 -6.47215 9.704E-27 2.746E-25 ZNF525 -89.533 -6.484 -6.40713 2.578E-38 1.317E-36 ZNF808 -86.94 -6.442 -6.30421 1.093E-21 2.274E-20 CRIP1 -80.273 -6.327 -6.30144 1E-109 2.68E-107 ZNF211 -80.046 -6.323 -6.1815 7.065E-21 1.405E-19 MYBPH -44.517 -5.476 -5.33045 9.951E-18 1.54E-16 AKT3 -36.41 -5.186 -5.1053 2.808E-29 9.224E-28 WWC3 -36.07 -5.173 -5.12008 2.41E-44 1.6E-42 ZNF888 -32.175 -5.008 -4.94001 9.888E-34 3.939E-32 ZNF816 -21.582 -4.432 -4.3169 1.894E-18 3.153E-17 PBX1 -12.275 -3.618 -3.60778 2.41E-158 1.52E-155 ZNF320 -11.371 -3.507 -3.48862 1.135E-82 2.053E-80 LRIG1 -10.133 -3.341 -3.29804 1.676E-35 7.226E-34 MMP14 -8.7061 -3.122 -3.09928 2.554E-61 2.679E-59 NR5A2 -7.688 -2.943 -2.93066 1E-103 2.42E-101 HKDC1 -7.6115 -2.928 -2.8542 9.208E-19 1.568E-17 DCLK1 -6.2159 -2.636 -2.59153 2.052E-27 6.033E-26 SLC1A3 -6.193 -2.63 -2.6218 1.4E-127 5E-125 ENKD1 -5.8638 -2.552 -2.49438 8.501E-21 1.684E-19 GGT5 -5.7985 -2.536 -2.43366 1.826E-12 1.819E-11 PRKAA2 -5.5329 -2.468 -2.45032 4.78E-61 4.884E-59

Table 18. Down-regulated gene sets enriched in U-87 MG GSTO1 KO RNA-seq (GSEA)

geneset size ES NES pValue fwer fdr

194 -0.66 -2.25 0 0 0 HALLMARK_E2F_TARGETS

HALLMARK_G2M_CHECKPOINT 193 -0.64 -2.17 0 0 0

HALLMARK_MYC_TARGETS_V1 195 -0.64 -2.16 0 0 0

HALLMARK_MYC_TARGETS_V2 58 -0.65 -1.84 0.00091 0.0104 0.003

KEGG_DNA_REPLICATION 36 -0.73 -1.91 0.00045 0.0322 0.041

HALLMARK_MITOTIC_SPINDLE 199 -0.44 -1.51 0.00333 0.277 0.077

HALLMARK_EPITHELIAL_MESENCHYMAL_ 194 -0.43 -1.46 0.00755 0.3787 0.094 TRANSITION

Table 19. Down-regulated gene sets enriched in HCT116 GSTO1 KO RNA-seq (GSEA)

geneset size ES NES pValue fwer fdr

GO_MITOCHONDRIAL_TRANSLATION 104 -0.69 -2.14 0 0.021 0.018

GO_TRANSLATIONAL_TERMINATION 90 -0.69 -2.11 0 0.0532 0.024

GO_SISTER_CHROMATID_SEGREGATION 161 -0.62 -2.06 0 0.1546 0.048

Table 20. Top 10 up-regulated gene sets enriched in U-87 MG GSTO1 KO RNA-seq (GSEA)

NE geneset size ES pValue fwer S

HALLMARK_BILE_ACID_METABOLISM 97 0.61 1.84 0.00017 0.0055

HALLMARK_KRAS_SIGNALING_DN 130 0.6 1.86 0 0.0036

KEGG_TASTE_TRANSDUCTION 33 0.76 1.91 0.00018 0.0099

GO_COMPLEMENT_ACTIVATION 25 0.81 1.93 0 0.1241

GO_MACROPHAGE_ACTIVATION 26 0.79 1.9 0.00036 0.2118

GO_DIGESTIVE_SYSTEM_PROCESS 43 0.73 1.93 0.00018 0.118

GO_PROTEIN_ACTIVATION_CASCADE 39 0.75 1.95 0 0.0721

GO_ASTROCYTE_DEVELOPMENT 16 0.86 1.86 0.00037 0.4121

KEGG_COMPLEMENT_AND_COAGULATION 49 0.66 1.76 0.0021 0.1519 _CASCADES

GO_NEURAL_CREST_CELL_MIGRATION 42 0.71 1.87 0.00018 0.3871

GO_REGULATION_OF_INTERLEUKIN_17_P 17 0.82 1.82 0.00092 0.6543 RODUCTION

Table 21. Top 10 up-regulated gene sets enriched in HCT116 GSTO1 KO RNA-seq (GSEA)

geneset size ES NES pValue fwer fdr

GO_RESPONSE_TO_INTERFERON_GAMMA 106 0.76 2.35 0 0 0

GO_RESPONSE_TO_TYPE_I_INTERFERON 52 0.87 2.41 0 0 0

GO_CELLULAR_RESPONSE_TO_INTERFERON_G 87 0.79 2.36 0 0 0 AMMA

GO_INTERFERON_GAMMA_MEDIATED_SIGNALIN 58 0.83 2.33 0 0 0 G_PATHWAY

GO_INNATE_IMMUNE_RESPONSE 415 0.66 2.33 0 0 0

GO_DEFENSE_RESPONSE_TO_VIRUS 128 0.74 2.33 0 0 0

GO_NEGATIVE_REGULATION_OF_VIRAL_PROCE 77 0.79 2.33 0 0 0 SS

GO_NEGATIVE_REGULATION_OF_MULTI_ORGA 124 0.74 2.32 0 0 0 NISM_PROCESS

GO_NEGATIVE_REGULATION_OF_VIRAL_GENO 44 0.84 2.28 0 0 0 ME_REPLICATION

GO_RESPONSE_TO_VIRUS 200 0.67 2.23 0 0 0

Table 22. Gene sets coexpressed with GSTO1 in glioblastoma (GBMLGG), gene size > 100

Geneset Size ES NES pValue fwer fdr

KEGG_OXIDATIVE_PHOSPHORYLATION 112 0.787706 3.797312 0 0.00E+00 0 KEGG_PARKINSONS_DISEASE 111 0.745272 3.578042 0 0.00E+00 0 KEGG_ALZHEIMERS_DISEASE 154 0.624366 3.162947 0 0.00E+00 0 KEGG_HUNTINGTONS_DISEASE 168 0.596927 3.054277 0 0.00E+00 0 KEGG_CYTOKINE_CYTOKINE_RECEPTOR_INTERAC 210 0.488027 2.581178 0 0.00E+00 0 TION KEGG_SYSTEMIC_LUPUS_ERYTHEMATOSUS 100 0.537534 2.53828 0 0.00E+00 0 HALLMARK_OXIDATIVE_PHOSPHORYLATION 200 0.689605 3.620629 0 0.00E+00 0 HALLMARK_ALLOGRAFT_REJECTION 188 0.591439 3.070587 0 0.00E+00 0 HALLMARK_INTERFERON_GAMMA_RESPONSE 199 0.555071 2.911628 0 0.00E+00 0 HALLMARK_INFLAMMATORY_RESPONSE 196 0.517923 2.708851 0 0.00E+00 0 HALLMARK_EPITHELIAL_MESENCHYMAL_TRANSITI 199 0.503027 2.642545 0 0.00E+00 0 ON HALLMARK_TNFA_SIGNALING_VIA_NFKB 198 0.494262 2.582616 0 0.00E+00 0 HALLMARK_COAGULATION 122 0.510019 2.485904 0 0.00E+00 0 HALLMARK_FATTY_ACID_METABOLISM 151 0.477637 2.413199 0 0.00E+00 0 HALLMARK_COMPLEMENT 190 0.452846 2.362397 0 0.00E+00 0 HALLMARK_MTORC1_SIGNALING 200 0.447988 2.355013 0 0.00E+00 0 HALLMARK_MYC_TARGETS_V1 200 0.446877 2.352636 0 0.00E+00 0 HALLMARK_GLYCOLYSIS 195 0.446602 2.341281 0 0.00E+00 0 HALLMARK_APOPTOSIS 159 0.451689 2.295267 0 0.00E+00 0 HALLMARK_IL2_STAT5_SIGNALING 198 0.414039 2.172424 0 6.40E-06 1.0E-4 HALLMARK_P53_PATHWAY 197 0.415461 2.173453 0 6.72E-06 1.0E-4 HALLMARK_HYPOXIA 196 0.423057 2.217768 0 7.08E-06 1.0E-4 HALLMARK_DNA_REPAIR 150 0.440402 2.218453 0 7.47E-06 1.0E-4 HALLMARK_ADIPOGENESIS 196 0.430889 2.251089 0 7.91E-06 1.0E-4 KEGG_LYSOSOME 118 0.498183 2.423058 0 8.70E-06 1.0E-4 HALLMARK_XENOBIOTIC_METABOLISM 183 0.392959 2.037972 0 2.64E-05 4.0E-4 HALLMARK_UV_RESPONSE_UP 157 0.391473 1.987984 0 5.81E-05 9.0E-4 HALLMARK_KRAS_SIGNALING_UP 195 0.357063 1.865255 0 2.80E-04 0.0043 HALLMARK_UNFOLDED_PROTEIN_RESPONSE 113 0.358447 1.728248 1.96E-4 1.49E-03 0.0241

Table 23. Gene sets coexpressed with GSTO1 in kidney renal clear cell carcinoma (KIRC), gene size > 100

Geneset Size ES NES pValue fwer fdr

KEGG_OXIDATIVE_PHOSPHORYLATION 111 0.802461 3.68746 0 0 0

KEGG_PARKINSONS_DISEASE 109 0.745635 3.385453 0 0 0

KEGG_ALZHEIMERS_DISEASE 154 0.62189 3.001112 0 0 0

KEGG_HUNTINGTONS_DISEASE 166 0.588283 2.859372 0 0 0

HALLMARK_OXIDATIVE_PHOSPHORYLATION 200 0.711186 3.542674 0 0 0

HALLMARK_MYC_TARGETS_V1 200 0.490823 2.44689 0 0 0

HALLMARK_ADIPOGENESIS 196 0.467361 2.323614 0 0 0

HALLMARK_FATTY_ACID_METABOLISM 154 0.480897 2.309315 0 0 0

HALLMARK_DNA_REPAIR 150 0.471583 2.250109 0 0 0

HALLMARK_MTORC1_SIGNALING 200 0.375916 1.872733 0 5.306162E-4 0.003

HALLMARK_XENOBIOTIC_METABOLISM 194 0.374869 1.85886 0 5.4909114E-4 0.0035

HALLMARK_PEROXISOME 101 0.38425 1.731763 0 0.002188806 0.0155

HALLMARK_GLYCOLYSIS 198 0.342799 1.704505 0 0.002579171 0.0202 HALLMARK_UV_RESPONSE_UP 153 0.350099 1.6858 0 0.002868925 0.0242

Table 24. Gene sets coexpressed with GSTO1 in skin cutaneous melanoma (SKCM), gene size > 100

Geneset Size ES NES pValue fwer fdr

KEGG_OXIDATIVE_PHOSPHORYLATION 111 0.785789 4.009629 0 0 0

KEGG_PARKINSONS_DISEASE 109 0.681525 3.451552 0 0 0

KEGG_LYSOSOME 120 0.607264 3.127597 0 0 0

KEGG_ALZHEIMERS_DISEASE 149 0.573336 3.047353 0 0 0

KEGG_HUNTINGTONS_DISEASE 165 0.519446 2.806686 0 0 0

HALLMARK_OXIDATIVE_PHOSPHORYLATION 200 0.682942 3.78869 0 0 0

HALLMARK_FATTY_ACID_METABOLISM 150 0.461695 2.466423 0 0 0

HALLMARK_ADIPOGENESIS 195 0.413927 2.296737 0 6.89E-05 1.0E-4

HALLMARK_DNA_REPAIR 150 0.396878 2.122251 0 9.35E-05 2.0E-4

HALLMARK_MTORC1_SIGNALING 200 0.386458 2.151331 0 1.09E-04 2.0E-4

HALLMARK_XENOBIOTIC_METABOLISM 180 0.336008 1.851719 0 8.95E-04 0.0033

HALLMARK_UV_RESPONSE_UP 156 0.34929 1.860824 0 9.06E-04 0.003

HALLMARK_MYC_TARGETS_V1 200 0.327569 1.822294 0 0.001083559 0.0045

HALLMARK_GLYCOLYSIS 197 0.321303 1.78311 0 0.001473649 0.0066

Table 25. Gene sets coexpressed with GSTO1 in stomach adenocarcinoma (STAD), gene size > 100

Geneset Size ES NES pValue fwer fdr

KEGG_OXIDATIVE_PHOSPHORYLATION 111 0.800803 3.664717 0 0 0

KEGG_PARKINSONS_DISEASE 109 0.780983 3.550849 0 0 0

KEGG_HUNTINGTONS_DISEASE 164 0.646332 3.135813 0 0 0

KEGG_ALZHEIMERS_DISEASE 152 0.6228 2.990071 0 0 0

KEGG_SPLICEOSOME 126 0.603232 2.802602 0 0 0

HALLMARK_MYC_TARGETS_V1 200 0.767117 3.818957 0 0 0

HALLMARK_OXIDATIVE_PHOSPHORYLATION 200 0.752524 3.747622 0 0 0

HALLMARK_INTERFERON_GAMMA_RESPONSE 200 0.596177 2.967457 0 0 0

HALLMARK_ALLOGRAFT_REJECTION 196 0.594643 2.945341 0 0 0

HALLMARK_MTORC1_SIGNALING 200 0.565942 2.818265 0 0 0

HALLMARK_E2F_TARGETS 200 0.549798 2.737318 0 0 0

HALLMARK_DNA_REPAIR 150 0.546395 2.619894 0 0 0

HALLMARK_FATTY_ACID_METABOLISM 152 0.492478 2.354612 0 0 0

HALLMARK_ADIPOGENESIS 197 0.461145 2.290671 0 0 0

KEGG_SYSTEMIC_LUPUS_ERYTHEMATOSUS 120 0.478062 2.216213 0 8.02E-05 0.001

HALLMARK_COMPLEMENT 198 0.38784 1.930437 0 2.69E-04 0.0022

HALLMARK_INFLAMMATORY_RESPONSE 199 0.378542 1.888832 0 2.99E-04 0.0029

2.151926 HALLMARK_UNFOLDED_PROTEIN_RESPONSE 113 0.413542 1.893205 3.10E-04 0.0028 E-4

HALLMARK_APOPTOSIS 160 0.371861 1.796382 0 7.88E-04 0.0086

HALLMARK_UV_RESPONSE_UP 157 0.372335 1.800518 0 8.11E-04 0.0083

HALLMARK_XENOBIOTIC_METABOLISM 191 0.350547 1.733989 0 0.001482781 0.0182

HALLMARK_G2M_CHECKPOINT 200 0.344341 1.713366 0 0.001871697 0.0238

Table 26. Gene sets coexpressed with GSTO1 in uveal melanoma (UVM), gene size > 100

Geneset Size ES NES pValue fwer fdr

KEGG_PARKINSONS_DISEASE 109 0.640309 2.950932 0 0 0

KEGG_OXIDATIVE_PHOSPHORYLATION 112 0.632242 2.928861 0 0 0

KEGG_ALZHEIMERS_DISEASE 150 0.52488 2.546679 0 0 0

KEGG_CYTOKINE_CYTOKINE_RECEPTOR_INTER ACTION 190 0.475214 2.383843 0 0 0

HALLMARK_OXIDATIVE_PHOSPHORYLATION 200 0.579882 2.929479 0 0 0

HALLMARK_ALLOGRAFT_REJECTION 181 0.511451 2.548116 0 0 0

HALLMARK_INTERFERON_GAMMA_RESPONSE 198 0.474115 2.387778 0 0 0

HALLMARK_MTORC1_SIGNALING 200 0.442105 2.233932 0 0 0

KEGG_HUNTINGTONS_DISEASE 164 0.459965 2.25131 0 1.19E-05 1.0E-4

HALLMARK_INFLAMMATORY_RESPONSE 184 0.40245 2.015198 0 1.32E-04 9.0E-4

HALLMARK_TNFA_SIGNALING_VIA_NFKB 193 0.403162 2.024733 0 1.51E-04 9.0E-4

KEGG_NATURAL_KILLER_CELL_MEDIATED_CYT OTOXICITY 101 0.453327 2.063369 0 4.96E-04 0.0047

HALLMARK_COMPLEMENT 182 0.371131 1.848579 0 8.71E-04 0.0073

HALLMARK_GLYCOLYSIS 193 0.357687 1.794317 0 0.001520485 0.014

1.466275 HALLMARK_FATTY_ACID_METABOLISM 150 0.358566 1.736468 7E-4 0.002554637 0.0252

1.434926 HALLMARK_IL2_STAT5_SIGNALING 195 0.342876 1.725465 1E-4 0.002775487 0.0296 4.439840 HALLMARK_APOPTOSIS 156 0.3465 1.690108 0.003871657 0.0444 3E-4

Table 27. Top 30 gene sets enriched in U-87 MG GSTO1 KO RNA-Seq (DAVID)

geneset pval qval num total GO:0030335~positive regulation of cell migration 2.96E-09 2.97E-06 85 393 GO:0040017~positive regulation of locomotion 3.47E-09 3.13E-06 89 420 GO:0051272~positive regulation of cellular component 5.98E-09 4.49E-06 88 418 movement GO:2000147~positive regulation of cell motility 7.56E-09 5.25E-06 86 407 GO:0048514~blood vessel morphogenesis 2.42E-08 1.29E-05 97 490 GO:0001525~angiogenesis 6.16E-08 2.78E-05 84 412 GO:0030198~extracellular matrix organization 1.2E-06 0.000308 68 333 GO:0043062~extracellular structure organization 1.34E-06 0.000317 68 334 GO:1904018~positive regulation of vasculature 1.47E-05 0.002289 35 143 development GO:0030278~regulation of ossification 1.76E-05 0.002484 41 181 GO:0032963~collagen metabolic process 2.17E-05 0.002791 29 110 GO:0001503~ossification 2.9E-05 0.003434 67 358 hsa04142:Lysosome 2.73E-05 0.003895 31 121 GO:1903555~regulation of tumor necrosis factor 4.26E-05 0.004457 28 108 superfamily cytokine production GO:0044259~multicellular organismal macromolecule 5.16E-05 0.005222 29 115 metabolic process GO:0071496~cellular response to external stimulus 5.25E-05 0.005253 51 255 GO:0014032~neural crest cell development 5.51E-05 0.005394 21 70 GO:0031589~cell-substrate adhesion 6.12E-05 0.005792 58 304 GO:0048762~mesenchymal cell differentiation 6.29E-05 0.005894 39 178 GO:0001655~urogenital system development 6.85E-05 0.006286 60 319 GO:0071706~tumor necrosis factor superfamily cytokine 7.1E-05 0.006453 28 111 production GO:0001819~positive regulation of cytokine production 7.52E-05 0.006698 69 383 GO:1904035~regulation of epithelial cell apoptotic 7.51E-05 0.006751 20 66 process GO:0014031~mesenchymal cell development 7.85E-05 0.006922 37 167 GO:0014065~phosphatidylinositol 3-kinase signaling 8.07E-05 0.007041 26 100 GO:0035455~response to interferon-alpha 8.75E-05 0.007561 10 19 hsa04512:ECM- interaction 2.68E-05 0.007634 25 87 GO:0014033~neural crest cell differentiation 9.41E-05 0.007835 22 78 GO:0060485~mesenchyme development 9.64E-05 0.007952 47 234 GO:0050920~regulation of chemotaxis 0.000132 0.010009 39 184

Table 28. Top 30 gene sets enriched in HCT116 GSTO1 KO RNA-Seq (DAVID)

geneset pval qval num total GO:0034340~response to type I interferon 1.2E-13 1E-09 31 77 GO:0060337~type I interferon signaling pathway 8.6E-12 1.79E-08 28 73 GO:0071357~cellular response to type I interferon 8.6E-12 1.79E-08 28 73 GO:0009615~response to virus 5.45E-09 2.07E-06 58 307 GO:0060333~interferon-gamma-mediated 1.95E-08 6.51E-06 24 75 signaling pathway GO:0045071~negative regulation of viral genome 4.2E-08 1.25E-05 19 50 replication hsa04612:Antigen processing and presentation 6.62E-08 1.85E-05 23 71 GO:0045069~regulation of viral genome replication 7.46E-08 1.95E-05 24 80 GO:0051607~defense response to virus 7.25E-08 1.95E-05 45 226 GO:1903901~negative regulation of viral life cycle 9.33E-08 2.29E-05 25 87 GO:0048525~negative regulation of viral process 9.27E-08 2.35E-05 28 106 GO:0048514~blood vessel morphogenesis 1.37E-07 3.09E-05 76 490 GO:0002483~antigen processing and presentation 2.45E-07 4.99E-05 10 14 of endogenous peptide antigen GO:0022407~regulation of cell-cell adhesion 2.7E-07 5.24E-05 62 376 GO:0034341~response to interferon-gamma 3.14E-07 5.83E-05 34 154 GO:0071346~cellular response to interferon- 4.73E-07 7.9E-05 30 128 gamma GO:0050792~regulation of viral process 6.08E-07 9.23E-05 47 259 GO:0050680~negative regulation of epithelial cell 9.48E-07 0.000136 28 118 proliferation GO:0050865~regulation of cell activation 1.09E-06 0.000149 71 471 GO:0019079~viral genome replication 1.08E-06 0.000151 26 105 GO:0019883~antigen processing and presentation 1.2E-06 0.000154 10 16 of endogenous antigen GO:0050678~regulation of epithelial cell 1.43E-06 0.000171 51 300 proliferation GO:0001525~angiogenesis 1.41E-06 0.000175 64 412 GO:0019885~antigen processing and presentation 1.92E-06 0.000214 9 13 of endogenous peptide antigen via MHC class I GO:0043903~regulation of symbiosis, 2.93E-06 0.000288 48 282 encompassing mutualism through parasitism GO:0043901~negative regulation of multi- 3.34E-06 0.000313 32 155 organism process GO:0051249~regulation of lymphocyte activation 4.22E-06 0.000374 59 381 GO:0007067~mitotic nuclear division 4.41E-06 0.000384 64 426 GO:0050673~epithelial cell proliferation 4.7E-06 0.000404 55 347 GO:1903037~regulation of leukocyte cell-cell 4.76E-06 0.000406 50 304 adhesion

Table 29. Top 30 up-regulated in HCT116 GSTO1 KO Proteomics

gene Description fc log2fc qval protein kinase (cAMP-dependent, catalytic) inhibitor PKIB 4.043 2.02 0 beta(PKIB) DHRS2 dehydrogenase/reductase 2(DHRS2) 3.466 1.79 0 INPP4B inositol polyphosphate-4-phosphatase type II B(INPP4B) 2.87 1.52 0 VSNL1 visinin like 1(VSNL1) 2.676 1.42 0 SLC31A1 solute carrier family 31 member 1(SLC31A1) 2.446 1.29 0.026 DYNLT3 dynein light chain Tctex-type 3(DYNLT3) 2.377 1.25 0.001 MTA3 metastasis associated 1 family member 3(MTA3) 2.355 1.24 0 calcium/calmodulin dependent protein kinase II CAMK2D 2.224 1.15 0 delta(CAMK2D) SYK spleen associated tyrosine kinase(SYK) 2.089 1.06 0.006 STAMBPL1 STAM binding protein like 1(STAMBPL1) 1.982 0.99 0.003 FUT8 fucosyltransferase 8(FUT8) 1.876 0.91 0.001 MAOB monoamine oxidase B(MAOB) 1.755 0.81 0 CD109 CD109 molecule(CD109) 1.723 0.79 0 ELF1 E74 like ETS 1(ELF1) 1.701 0.77 0.001 FAS Fas cell surface death receptor(FAS) 1.688 0.76 0 S100A4 S100 calcium binding protein A4(S100A4) 1.677 0.75 0 TRIM72 Fas cell surface death receptor(FAS) 1.68 0.75 0 GSN (GSN) 1.672 0.74 0 TCF7L2 transcription factor 7 like 2(TCF7L2) 1.666 0.74 0 ACSL5 acyl-CoA synthetase long-chain family member 5(ACSL5) 1.622 0.7 0 TP53I3 tumor protein inducible protein 3(TP53I3) 1.618 0.69 0 MTUS1 associated tumor suppressor 1(MTUS1) 1.609 0.69 0 SAMD9 sterile alpha motif domain containing 9(SAMD9) 1.583 0.66 0 PDLIM5 microtubule associated tumor suppressor 1(MTUS1) 1.583 0.66 0 OSBPL7 oxysterol binding protein like 7(OSBPL7) 1.579 0.66 0.004 ARHGEF10L Rho guanine nucleotide exchange factor 10 like(ARHGEF10L) 1.58 0.66 0.005 SERPINB5 serpin family B member 5(SERPINB5) 1.555 0.64 0 GALNT5 polypeptide N-acetylgalactosaminyltransferase 5(GALNT5) 1.561 0.64 0.005

Table 30. Top 30 down-regulated proteins in HCT116 GSTO1 KO Proteomics

gene fc log2fc qval CRIP1 cysteine rich protein 1 -62.5 -5.94 0.003 MICALL1 MICAL like 1 -23.2558 -4.53 0 GSTO1 glutathione S-transferase omega 1 -18.8679 -4.24 0 BAX BCL2 associated X, apoptosis regulator -15.625 -3.97 0.001 STXBP6 syntaxin binding protein 6 -7.69231 -2.94 0.006 IFRD2 interferon related developmental regulator 2 -7.63359 -2.93 0.039 SEC31A SEC31 homolog A, COPII coat complex component -5.43478 -2.44 0 HMGN5 high mobility group nucleosome binding domain 5 -5.43478 -2.44 0.001 MVK mevalonate kinase -3.83142 -1.94 0.005 F3 coagulation factor III, tissue factor(F3) -3.11526 -1.64 0.055 SH3 domain binding glutamate rich protein SH3BGRL -2.71739 -1.44 0 like(SH3BGRL) SUSD2 sushi domain containing 2(SUSD2) -2.54453 -1.35 0 SRGAP1 SLIT-ROBO Rho GTPase activating protein 1(SRGAP1) -2.33645 -1.22 0 TAGLN transgelin(TAGLN) -2.331 -1.22 0.005 TGM2 transglutaminase 2(TGM2) -2.23714 -1.16 0 SCAF1 SR-related CTD associated factor 1(SCAF1) -2.07469 -1.05 0 SLC2A1 solute carrier family 2 member 1(SLC2A1) -2.06186 -1.04 0 methylmalonic aciduria (cobalamin deficiency) cblA MMAA -2.06186 -1.04 0.093 type(MMAA) BST2 bone marrow stromal cell antigen 2(BST2) -1.9685 -0.98 0.004 SCD stearoyl-CoA desaturase(SCD) -1.95313 -0.96 0 STK39 serine/threonine kinase 39(STK39) -1.9305 -0.95 0 CA2 carbonic anhydrase 2(CA2) -1.90476 -0.93 0 MYL9 light chain 9(MYL9) -1.80832 -0.85 0 WDTC1 WD and tetratricopeptide repeats 1(WDTC1) -1.76056 -0.82 0.024 SIL1 SIL1 nucleotide exchange factor(SIL1) -1.72414 -0.78 0.002 PROSER2 proline and serine rich 2(PROSER2) -1.66945 -0.74 0 EAPP associated phosphoprotein(EAPP) -1.65563 -0.73 0.002 TNRC6C trinucleotide repeat containing 6C(TNRC6C) -1.65563 -0.73 0.005 DNMT1 DNA methyltransferase 1(DNMT1) -1.65289 -0.73 0 TCF20 transcription factor 20(TCF20) -1.63934 -0.71 0

Table 31. Up-regulated gene sets enriched in HCT116 GSTO1 KO Proteomics

Geneset Size ES NES pValue fwer fdr

HALLMARK_INTERFERON_ALPHA_RESPONSE 48 0.588564 2.004396 0 0.0012 0.001192

HALLMARK_INTERFERON_GAMMA_RESPONSE 84 0.494596 1.862089 0.000186 0.0136 0.006575

KEGG_ENDOMETRIAL_CANCER 33 0.551261 1.745032 0.00419 0.2578 0.044102

KEGG_ACUTE_MYELOID_LEUKEMIA 33 0.5478 1.728918 0.005667 0.2973 0.046894

KEGG_ALZHEIMERS_DISEASE 92 0.458554 1.750742 0.00056 0.2467 0.0471

HALLMARK_NOTCH_SIGNALING 16 0.625238 1.64775 0.018182 0.1364 0.047554

KEGG_TRYPTOPHAN_METABOLISM 18 0.647011 1.753232 0.004868 0.2417 0.052488

KEGG_GLIOMA 33 0.557694 1.762952 0.004435 0.2219 0.055791

KEGG_NATURAL_KILLER_CELL_MEDIATED_ 42 0.538937 1.779483 0.001821 0.1909 0.056004 CYTOTOXICITY

KEGG_PHOSPHATIDYLINOSITOL_SIGNALING_ 36 0.521571 1.66369 0.01114 0.4822 0.062116 SYSTEM

KEGG_PROSTATE_CANCER 43 0.498524 1.653768 0.00866 0.5148 0.06363

KEGG_MELANOMA 27 0.555522 1.667704 0.01444 0.4693 0.064578

KEGG_ARRHYTHMOGENIC_RIGHT_VENTRICULAR_ 22 0.585459 1.671777 0.013151 0.4567 0.067623 CARDIOMYOPATHY_ARVC

KEGG_VALINE_LEUCINE_AND_ISOLEUCINE_ 35 0.557598 1.781382 0.003271 0.1882 0.068953 DEGRADATION

KEGG_FATTY_ACID_METABOLISM 28 0.552116 1.674393 0.012219 0.4488 0.071988

KEGG_VIRAL_MYOCARDITIS 24 0.549899 1.604244 0.018013 0.6699 0.080675

KEGG_CELL_ADHESION_MOLECULES_CAMS 21 0.570603 1.609553 0.025972 0.6534 0.08166

KEGG_ERBB_SIGNALING_PATHWAY 50 0.471509 1.613452 0.008584 0.6407 0.084099

KEGG_THYROID_CANCER 19 0.647056 1.781913 0.006361 0.1877 0.091547

Table 32. Down-regulated gene sets enriched in HCT116 GSTO1 KO Proteomics

Geneset Size ES NES pValue fwer fdr

HALLMARK_CHOLESTEROL_HOMEOSTASIS 44 -0.63555 -2.04589 0.001957 0.041 0.042738