Supporting Information Microrna-202-3P Inhibits Cell

Wang, et al. miR-202-3p targets ARL5A in CRC

Supporting information

MicroRNA-202-3p Inhibits Cell Proliferation by Targeting ADP-Ribosylation Factor-Like 5A in Human Colorectal Carcinoma

*Qifeng Wang1,2,3,4, *Zhaohui Huang5, *Weijie Guo2,4,6, Shujuan Ni1,2,3,4, Xiuying Xiao8, Lisha Wang1,2,3,4, Dan Huang1,2,3,4, Cong Tan1,2,3,4, Qinghua Xu1,2,3,4,7, Ruopeng Zha2,4,6, JiweiZhang5, Weiqi Sheng1,2,3,4, Xianghuo He6 #, Xiang Du1,2,3,4 #

1Department of Pathology, Fudan University Shanghai Cancer Center, Shanghai, China 2Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China 3Institute of Pathology, Fudan University, Shanghai, China 4Institutes of Biomedical Sciences, Fudan University, Shanghai, China 5Wuxi Oncology Institute, the Affiliated Hospital of Jiangnan University, Wuxi, China 6State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai Jiao Tong University School of Medicine, Shanghai, China 7Fudan University Shanghai Cancer Center - Institute Mérieux Laboratory, Shanghai, China 8Department of Oncology, Ren Ji Hospital, Shanghai Jiao Tong University, Shanghai, China

*These authors contributed equally to this work.

# Correspondence:

Xiang Du, Department of Pathology, Fudan University Shanghai Cancer Center, 270 Dong An Road, Shanghai, 200032, China. Tel/Fax: 86-21-64170067; E-mail: [email protected]. 1

Wang, et al. miR-202-3p targets ARL5A in CRC

Xianghuo He, State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, No.25/Ln.2200, Xietu Road, Shanghai 200032, China.

Telephone /Fax: 86-21-64436539; E-mail: [email protected].

Wang, et al. miR-202-3p targets ARL5A in CRC

Supplementary Tables Table S1. Patient information.

Characteristics Colorectal cancer（n=94）

Gender Male 50 Female 44 Age at diagnosis Mean ± SD 60±11 Median(range) 57(31-82) TNM stage I 9 II 27 III 42 IV 16 Nodal status Positive 58 Negative 36 T stage T1 4 T2 14 T3 31 T4 45 Tumor location Rectum 40 Distal colon 25 Proximal colon 29 Histology Adenocarcinoma 90 Mucous adenocarcinoma 4 signet-ring cell carcinoma 0

Wang, et al. miR-202-3p targets ARL5A in CRC

Table S2. Primers sequences. Primer name Sequence(5' to 3') Product size bp

Primers for the detecting of pre-mir-202 copy number variation Reverse primer partially complementary with pre-mir-202 pre-mir-202-CNV-F1 CAGGTAAGACGAGGCCCAAG 206 pre-mir-202-CNV-R1 TAGGAAAAAGGAACGGCGGG Forward primers partially complementary with pre-mir-202 pre-mir-202-CNV-F2 TAGGGCATGGGAAAACGGG 274 pre-mir-202-CNV-R2 CAGAGATCAACCCAGGGCAG PCR products covers the full length of pre-mir-202 pre-mir-202-CNV-F3 TCAGTCCCGACTTCTCCCTG 306 pre-mir-202-CNV-R3 CCAAGGATGTGGGGGTCAAA

LINE-1-CNV-F AAAGCCGCTCAACTACATGG 122

LINE-1-CNV-R TGCTTTGAATGCGTCCCAGAG

Primers for the validation of gene expression C7orf58-RT-F AGCCACTTTGCAGAAAGGAA C7orf58-RT-R GACGTCACCAACACATACGC 255 CPEB4-RT-F TGCATTCCTGCTGTTTCAAG CPEB4-RT-R TATCATCGCAAGCTCCACAG 221 MOBKL3-RT-F AAGGCCAAGATGAAGGTGTG MOBKL3-RT-R TCTGGATGGCATTCACTCTG 267 HECTD2-V1-RT-F TCCTATCCAGCCCAAAACTG HECTD2-V1-RT-R ATGAGGCAGTGGCATCTTTC 215 HAS2-RT-F AAACAGTTGCCCTTTGCATC HAS2-RT-R ACGTGTTGCGAGCTTTCTTT 221 KIAA1609-RT-F CTGACTACCCTGGTCCCTGA KIAA1609-RT-R CTTCACCTCCCAAGAGCAAG 213 ARL5A-V3-RT-F AGCGCATGAGGACCTAAGAA ARL5A-V3-RT-R CAATCCCTCGCCAGTTAGAG 134 TSPAN2-RT-F TCCGAACAGGTCCAACCTAC TSPAN2-RT-R TGAGTTTCGTATCGCACAGC 192 LAMA1-RT-F AGCGGATATGCAGCTCTTGT LAMA1-RT-R GTTATCCTGCCAGCACCATT 195 RAG1-RT-F GCAGCTATTGTCCCTCTTGC RAG1-RT-R CATTGCACTCTTTTGCTGGA 169 BCL7A-RT-F TGGTGACACATCCCTACGAA BCL7A-RT-R ATGGAGCTCTGGTTGCTGTT 232

Wang, et al. miR-202-3p targets ARL5A in CRC

GALNT12-RT-F ATAACACCCACCAGCCTGAG GALNT12-RT-R TTCGATGGGCTCCTTGATAC 480 LRIG3-RT-F GCAGGATGGGTACGTGTCTT LRIG3-RT-R TTCCACATCAGCTTCACTGC 241 PTPRD-RT-F TTTACACGAACACCCGTTGA PTPRD-RT-R GAATTTGATCTTCCCGCAAA 201 RGS16-RT-F CAAGACACGTCTGGGGATCT RGS16-RT-R CAGGTCGAACGACTCTCTCC 141 TET2-RT-F TTTGCCAGCCTCCTTTTCTA TET2-RT-R ATGTGGGTGAAAATGGGAAA 165 PCTP-RT-F GAAGAGCACCCACCACTGTT PCTP-RT-R GACGAGTGAGCTGGATGTCA 173 TET3-RT-F CCAAGAGTCTGCTGGACACA TET3-RT-R CTCCTTCCCCGTGTAGATGA 162

Primers for the cloning of pre-miR-202 pre-miR-202-OUT-F CGGCTCAGAGAGTTCTCCAC 654 pre-miR-202-OUT-R CCACACAAAAGCACACATCC pre-miR-202-IN-F CCGCTCGAGTTGGCTCTGCCCTCCTCA 415 pre-miR-202-IN-R GCGACGCGTATCCACGTTTCCCTTGTTTT

Primers for the cloning of target gene 3'-UTR binding sites ARL5A-OUT-F AGCGCATGAGGACCTAAGAA ARL5A-OUT-R TCTGCTAGCTTTCCCACTCTG 982 ARL5A-IN-F CCGGAATTCAGTGCTGGACTTTACCTGAAAGC ARL5A-IN-R CGCGGATCCAGCTTTCCCACTCTGTTG 745 ARL5A-Mut-F AAGGTTGTGTGTGTATTGTGTATAATGGACAAGTTCAAGTT 326 ARL5A-Mut-R AATCAATGCCATTAACTTGAACTTGTCCATTATACACAATA 473 BCL7A-OUT-F GTTTTTGGGGGCTAATTGGT BCL7A-OUT-R CCTAGAGGAATCGGTCCACA 1135 BCL7A-IN-F CCGGAATTCGTGGCTCTTTCTCCTGACC BCL7A-IN-R TTCCGCTCGAGTCACCAAGATGGAGTTTCG 458 CPEB4-OUT1-F CATTCTGACCCCTTCCTCAA CPEB4-OUT1-R TTTCATGTTTTGCCCACTGA 1496 CPEB4-IN1-F CCGGAATTCAGTTACCACCACATCAGA CPEB4-IN1-R CGCGGATCCAGTTCAACAGTGCAAAAC 581 CPEB4-OUT2-F CACCTGGCTTATGACCCCTA CPEB4-OUT2-R GTGCTGGGGAAGGATGTTTA 808 CPEB4-IN2-F CCGGAATTCAAATGCTTGTAGGTGGCTTG CPEB4-IN2-R CGCGGATCCCTGGGGAAGGATGTTTAA 516

Wang, et al. miR-202-3p targets ARL5A in CRC

HECTD2-OUT-F TCTTTGAGGAAAAGGGGTCA HECTD2-OUT-R TTTGAAAAATTGCATCCAGAA 864 HECTD2-IN-F CCGGAATTCGAGATGACTCATGGAAAA HECTD2-IN-R CGCGGATCCAAAAGGAAGGACGTAAAA 517 KIAA1609-OUT-F CAGATGGGAAAGTTGGGAGA KIAA1609-OUT-R CACGCCCTAAAAAGGTTTGA 1268 KIAA1609-IN-F CGGGGTACCCAGATGGGAAAGTTGGGAGAT KIAA1609-IN-R TTCCGCTCGAGCAGGGCTATTGCTAAGTGGAAG 695 LRIG3-OUT-F GCAGGATGGGTACGTGTCTT LRIG3-OUT-R TCCATAAAAAGAAATCTGGCATT 1068 LRIG3-IN-F CCGGAATTCCGAACGGAGCTTCAGTAATATATC LRIG3-IN-R CGCGGATCCGCATCATTCCCAGTATAAAA 616 MOBKL3-OUT-F CCCAGCAGGGTTAGCATAAA MOBKL3-OUT-R CATCAACACCCTTGGAACCT 1362 MOBKL3-IN-F CCGGAATTCTTGCTGCTAAACAATCATTGG MOBKL3-IN-R CGCGGATCCAAACATAAACTTCGTGGG 802 PCTP-OUT-F AAGGCTGCCTGTACCTCTCA PCTP-OUT-R AGAACACACTTTTCTTCCCAGA 496 PCTP-IN-F CGGGGTACCACTGAGGCGTCAAGATGG PCTP-IN-R TTCCGCTCGAGCTTTTCTTCCCAGAAAGAA 468 TET2-OUT-F GCTCCAACTCATGGGTCAAT TET2-OUT-R CAGCAGATGCATGGCTAAAA 1472 TET2-IN-F CGGGGTACCCCAGACTATGTGCCTCAGA TET2-IN-R TTCCGCTCGAGCATATCATCTCGTCCAATAA 530 TET3-OUT1-F CAGCACAAGAACCTCAACCA TET3-OUT1-R CCATGACATCGTGTGACTCC 1167 TET3-IN1-F CCGGAATTCGGGAAGCCAAGATGAAGCAGC TET3-IN1-R CGCGGATCCCACCCGACAGCCGTAGTT 515 TET3-OUT2-F TGGAAGGTCCCGAGTGTAAC TET3-OUT2-R GAAACATTGGCCCTGCTAAA 914 TET3-IN2-F CCGGAATTCGGTCCCGAGTGTAACCCAGG TET3-IN2-R CGCGGATCCCTGCTGCGAAGCGATTTT 791 TET3-OUT3-F TTGCTGAGGTGCTATCAACG TET3-OUT3-R ATTTCTGTGCTGCGGTCTCT 1290 TET3-IN3-F CCGGAATTCGTCTTCTCAGGCTTGATGTG TET3-IN3-R CGCGGATCCTTTGGTCCAGGGTATGTT 828

Primers for the cloning of ARL5A ORF ARL5A-OUT-F GCTGGATAATGCAGGGAAAA ARL5A-OUT-R TGAGAAAGCAAACTGGAAGG 657

Wang, et al. miR-202-3p targets ARL5A in CRC

ARL5A-IN-F CCGGAATTCATGAATGAAGTTGTACATACATCT ARL5A-IN-R CGCGGATCCTCATCTAATCTTAAGTCGT 447 * Sequences underlined are sites for restriction enzyme.

Table S3. siRNA sequences for potential target genes Gene name Detail Sequence ARL5A Target sequence GTACATACATCTCCTACAA si-5’-3’ GUACAUACAUCUCCUACAA dTdT si-3’-5’ dTdT CAUGUAUGUAGAGGAUGUU LRIG3 Target sequence GGACCAGTCTCGGCAAATA si-5’-3’ GGACCAGUCUCGGCAAAUAdTdT si-3’-5’ dTdT CCUGGUCAGAGCCGUUUAU HECTD2 Target sequence GGCCAAATTTCCACTTTCA si-5’-3’ GGCCAAAUUUCCACUUUCA dTdT si-3’-5’ dTdT CCGGUUUAAAGGUGAAAGU CPEB4 Target sequence CCTGCTGCTTCGGCTAATA si-5’-3’ CCUGCUGCUUCGGCUAAUA dTdT si-3’-5’ dTdT GGACGACGAAGCCGAUUAU

Table S4. Downregulated genes in miR-202-3p-transfected LoVo cells. Fold change GeneSymbol 0.001708 MX1 0.013141 CCL5 0.016712 IFI27 0.017039 IFITM1 0.01913 C20orf69 /// DKFZP434B2016 /// LOC643670 /// LOC728105 /// LOC728323 /// PCMTD2 0.023592 IFI44L 0.024209 CLEC1A 0.024731 BST2 0.028332 OAS2 0.028716 RTP4 0.02943 MBD2 0.029534 XAF1 0.035945 HSH2D 0.037625 BMS1P5 0.039029 IFIH1 0.040614 LIN7A 0.040683 IFI6

Wang, et al. miR-202-3p targets ARL5A in CRC

0.041435 CFTR 0.041468 GLT8D2 0.042305 CYP19A1 0.048665 CCDC89 0.049116 WDR78 0.056856 PTPN22 0.060213 LOC100132832 /// LOC441259 /// LOC730324 /// PMS2L1 /// PMS2L2 /// PMS2L5 0.060551 CCL3 /// CCL3L1 /// CCL3L3 /// LOC728830 0.061656 IL28A /// IL28B 0.061973 LOC643308 /// RPL7 /// RPL7P12 /// RPL7P20 /// RPL7P23 /// RPL7P24 /// RPL7P26 /// RPL7P48 /// RPL7P8 0.062314 GIMAP2 0.062946 LOC285889 0.06344 HRH4 0.064248 IL28A 0.065051 C3orf15 0.065807 C6orf150 0.066661 APOL1 0.069673 FPR2 0.070062 CXCL11 0.07008 HLA-F 0.070192 KIAA0999 0.070262 NCRNA00171 0.07082 C1orf161 0.07127 MX2 0.072042 PRSS12 0.072781 IL2RB 0.073039 ACCN3 0.074287 C12orf69 0.075144 CALD1 0.07651 PRLR 0.077343 GABRR1 0.07752 CXCL10 0.078672 TBX18 0.079611 TEX9 0.079743 TTLL13 0.079789 KRT14 0.080878 ALDOB 0.080913 PHLDB2 0.081091 OAS1 0.081328 TMEM61 0.082192 CALB1

Wang, et al. miR-202-3p targets ARL5A in CRC

0.08221 LAMA1 0.082678 DMGDH 0.083011 APIP 0.083724 CD244 0.084156 HNF4A 0.08613 PARP14 0.086587 GBP1 0.087316 LOC93432 0.093252 TRIM58 0.093732 CDR1 0.094757 C1S 0.094937 USP18 0.095598 NFKB2 0.095814 TRBV27 0.096062 GCET2 0.099278 EPSTI1 0.099452 HIBADH 0.101666 BATF2 0.102442 PTPRD 0.102761 UBE2L6 0.103153 MAP3K15 0.104268 SCN10A 0.104843 LRRTM4 0.105083 LOC730034 0.107542 PALM2 /// PALM2-AKAP2 0.107633 GREM1 0.107834 C10orf79 0.107896 KSR2 0.108817 TNFRSF9 0.110389 VMA21 0.110759 RERE 0.110999 DDX58 0.111506 STAT1 0.111976 RALGPS1 0.112417 CTSS 0.11361 RSAD2 0.115718 SEMA3D 0.116519 MPP7 0.118102 ZNF467 0.119663 ERICH1 0.120126 CLDN11 0.120648 NEO1 0.12204 TRIM22

Wang, et al. miR-202-3p targets ARL5A in CRC

0.12222 ITGB8 0.122829 LOC100129597 0.122906 NLRP1 0.123783 ANKRD22 0.124349 SYN2 0.126596 ST3GAL6 0.126612 NCF2 0.128026 PLEKHG2 0.128257 PARP9 0.128722 FAM105B 0.128938 BCL2L13 0.129371 IFIT1 0.130359 LOC283856 0.130596 C15orf48 0.133209 OAS3 0.133308 IRF9 0.133359 SPATA16 0.135535 SRC 0.138256 ELP1P 0.140437 PDZD2 0.142178 MTMR7 0.142887 C14orf102 0.142936 IL29 0.144256 SEMA6B 0.144984 HSPB8 0.145959 IFIT3 0.147324 NIACR2 0.148543 DDX60L 0.149314 TCAM1 0.150705 HERC6 0.15402 IL7R 0.15603 CPEB4 0.156123 APOL6 0.156362 OLFML2A 0.156799 PDZRN3 0.15704 PRIC285 0.157217 C1orf96 0.157653 OFCC1 0.1579 LOC100130429 0.159449 SBK1 0.159824 ISM1 0.161564 CEACAM1 0.162039 SPAG6

Wang, et al. miR-202-3p targets ARL5A in CRC

0.163625 EPHA4 0.166118 TSGA10 0.166176 PLSCR1 0.168007 GBP5 0.168988 ZCCHC2 0.170441 COMMD6 0.170595 SPEG 0.170684 SP2 0.172811 C21orf37 0.173058 RARRES3 0.173845 KIAA1654 0.174546 SNPH 0.176516 NTRK2 0.177324 CDH26 0.177801 FCER2 0.178592 TMEM88 0.179458 IL11 0.179559 ANO3 0.181421 SAMD9L 0.182836 C19orf66 0.184736 C8orf79 0.185133 BIRC3 0.185171 CDH1 0.185193 PSEN1 0.186453 OASL 0.186731 PSMB9 0.186875 SAMD9 0.188163 SGCZ 0.188169 RBM15 0.188327 C7orf58 0.189239 HEPH 0.191009 IFI44 0.191904 LRRC63 0.193673 ABCC11 0.194371 HERC5 0.19469 GRIK2 0.196787 USP44 0.19725 SLC17A4 0.197395 FAM118B 0.197622 HKDC1 0.198783 TAP1 0.199425 SVEP1 0.19956 IFIT2

Wang, et al. miR-202-3p targets ARL5A in CRC

0.200794 FMO2 0.201985 DDX60 0.202478 ACVR1C 0.204356 IFI35 0.204822 MFSD2 0.20582 ADAMDEC1 0.209946 TRIM73 /// TRIM74 0.212088 CNTNAP2 0.213971 DLEU2 0.2154 FRMD4B 0.217721 SP110 0.218404 DTX3L 0.218689 TMEM140 0.21935 SEC16B 0.220014 ABCC13 0.222555 ZNF225 0.222596 ZC3HAV1 0.223753 CTSLL3 0.223951 ISG20 0.224458 COL27A1 0.225738 PCSK6 0.226269 C4orf23 0.226708 ARL5A 0.226767 LOC93463 0.228868 POLR3G 0.228915 HIPK3 0.229022 ADRA1D 0.230842 PEX16 0.23145 PER3 0.232341 SERPINB11 0.233131 CASD1 0.2338 FBXO32 0.235073 DLGAP1 /// LOC388458 0.236582 C1orf38 0.237414 CASP1 0.237466 HOMEZ 0.237622 WIPI2 0.238658 CHM 0.238758 IRF7 0.239221 OGDH 0.241108 MAP3K8 0.24174 RP9P 0.241797 NR5A2

Wang, et al. miR-202-3p targets ARL5A in CRC

0.242198 LOC400940 0.242702 XG 0.243297 TNFSF10 0.244165 C9orf24 0.246312 SLC22A3 0.247414 ATP5SL 0.247968 LOC400796 0.24867 TDRD6 0.248883 CDH2 0.249289 VTI1A 0.250516 RGS16 0.250962 HNMT 0.251371 SAMD5 0.252196 RBMY2FP 0.252884 LOC100128718 0.253101 NBLA00301 0.255503 LOC120376 0.256984 CLIC3 0.257642 CD80 0.258195 IFI16 0.258629 TRPC4 0.258683 EPHB2 0.262667 GSTT1 0.26338 BDNFOS 0.264362 C1orf116 0.264691 PDE9A 0.264726 C21orf15 0.265745 RSU1 0.265915 NT5DC4 0.266535 PLCG2 0.270648 NFRKB 0.273322 NDUFA5 0.273504 IFNGR1 0.275385 ERG 0.275876 ATP5G3 0.276254 ITGB2 0.276559 THBS1 0.277181 PCOLCE2 0.277415 CIAPIN1 0.278511 SLC27A3 0.279647 SEMA3A 0.280023 CBLN2 0.280111 LRRC40

Wang, et al. miR-202-3p targets ARL5A in CRC

0.281604 CNKSR2 0.28202 RFX4 0.284022 WSB1 0.284134 RBPJL 0.284436 MBD5 0.284506 FGFBP2 0.285505 ZDHHC4 0.286804 HSF1 0.288393 MBP 0.288731 TRIM14 0.289432 C3 0.291416 OR51B6 0.293236 MAL2 0.294055 IFIT5 0.294083 LOC100133229 0.29564 LZTS1 0.296086 PTGFR 0.297079 BTN3A2 0.297238 LY6E 0.298333 PCDH7 0.29847 RAB11FIP4 0.298579 LOC653506 /// METRNL 0.298598 HOXB7 0.299063 IL23A 0.299725 CD24 0.300122 ARHGEF7 0.300188 NELL2 0.300219 PLEKHO1 0.301033 FAM46A 0.302728 CYC1 0.302879 TTTY9A /// TTTY9B 0.303188 HLA-C 0.303563 GPR155 0.305033 MRPL50 0.305169 VASH2 0.305459 RASGRP3 0.30653 FAM95B1 /// LOC100132340 /// LOC644587 /// LOC90499 0.307365 NT5C3L 0.307809 TNFAIP3 0.308359 C2 /// CFB 0.30923 LOC202181 0.309526 MMP13 0.309923 RAB15

Wang, et al. miR-202-3p targets ARL5A in CRC

0.310502 IGFBP6 0.310747 CIB4 0.311321 ZNF114 0.311668 SP100 0.311955 TMX3 0.311969 PTX3 0.312404 PPP2R2C 0.312495 HBP1 0.312769 TEX15 0.315179 TRIM21 0.316502 WBSCR23 0.318615 MSH4 0.318821 KIF13A 0.319748 UBR2 0.321664 SEMA4A 0.322178 CADM4 0.322579 IL6 0.323014 AS3MT 0.32325 TTBK2 0.323326 ELAVL4 0.323606 SOD2 0.324671 KIAA1211 0.324814 BTN3A3 0.324825 MGC57346 0.324943 SYNPO2 0.325232 ST8SIA4 0.325602 PARP12 0.325881 GDPD1 0.32613 FLT3LG 0.327812 APC2 0.327969 SCN3A 0.328178 FAM3C 0.328292 EIF2AK2 0.328371 ITGA2B 0.330727 JUNB 0.331381 SLC31A2 0.332287 AIM2 0.332587 C6orf211 0.334563 CHST11 0.335625 SPARC 0.336412 SIRT5 0.336664 TK2 0.338533 C6orf58

Wang, et al. miR-202-3p targets ARL5A in CRC

0.339569 TTC28 0.341097 LOC151162 /// MGAT5 0.342871 STAT6 0.343053 STAT2 0.344032 KCTD20 0.344641 PML 0.344655 HRASLS2 0.344915 SKAP2 0.345658 OTUD7B 0.346963 ERAP1 0.347245 C5orf56 0.347594 SSTR1 0.34772 SOCS3 0.349512 LAMP3 0.349688 FGF14 0.350333 ADIPOQ 0.350459 PATL2 0.351446 SLC25A23 0.351669 LOC100130476 0.352344 FUT4 0.352863 PSMB8 0.353161 PRRX1 0.353221 NNMT 0.353522 LOC493754 0.354473 ZNF33A 0.354678 GPR4 0.355008 AGXT2L1 0.35556 ZNRF1 0.355702 TDRD7 0.355907 VPREB1 0.356426 TLR3 0.356553 MDM2 0.356605 CSF2 0.357609 FAM66C /// FAM66D 0.359701 LOC100128439 0.359914 TSPAN12 0.36043 MTMR11 0.361677 SLC2A13 0.362023 HLA-B 0.362813 DLL3 0.363935 PCGF5 0.364003 SLC35D3 0.364453 TSPAN9

Wang, et al. miR-202-3p targets ARL5A in CRC

0.365041 TXNDC11 0.365289 PIN1L 0.365871 LYZL6 0.366184 LILRA1 0.366295 MCPH1 0.366649 DNAH14 0.366936 MEGF6 0.367164 KCNQ4 0.367273 MAN2A2 0.367428 ALS2CR11 0.367764 SAMHD1 0.368241 NCALD 0.368706 TET3 0.368795 CXCL1 0.368899 ZHX1 0.369665 HLA-E 0.369897 NSMCE4A 0.370124 CYLD 0.370611 PDGFRL 0.37191 ASTN1 0.372169 HAS2 0.372861 MCL1 0.373198 HS6ST3 0.373512 GPATCH4 0.373656 LOC285628 0.374104 PHF11 0.374119 INHBA 0.374743 ALS2 0.375955 MOBKL3 0.376172 ABCC9 0.376184 RBM43 0.378122 EML1 0.378334 BDNF 0.378369 ZNF230 0.378691 FGD4 0.37973 KLHL24 0.380132 RAD54L2 0.380239 CAPN9 0.380588 FGF18 0.380893 IFITM3 0.381305 CH25H 0.381858 CALN1 0.381891 AFF4

Wang, et al. miR-202-3p targets ARL5A in CRC

0.382497 HIST1H3J 0.382623 SAMD10 0.383392 PIGH 0.383439 HLA-G 0.38351 CHD1L 0.38368 RBM33 0.384388 CASP9 0.384508 CRLS1 0.385303 SLC5A3 0.386093 HIST1H3F 0.386696 IL6R 0.387597 GATM 0.387888 COL5A1 0.388208 SH3GLB1 0.389089 DUOX1 0.389594 C5orf13 0.389787 TCF4 0.391093 CASZ1 0.391229 VEPH1 0.391587 ICAM1 0.391826 KLF11 0.392695 FST 0.393236 LOC100130958 0.393647 ZNF620 0.394291 C9orf95 0.395308 TTC23 0.395642 FBXL17 0.395731 GUCA1A 0.397326 LOC100133308 0.3977 BLZF1 0.397731 CCL20 0.397757 RASEF 0.398187 STK3 0.398337 NRP2 0.39868 SERPINE1 0.398762 OR2A20P /// OR2A5 /// OR2A9P 0.399361 HIVEP2 0.39946 SLC6A19 0.399473 LOC653562 /// SLC6A10P /// SLC6A8 0.399551 IKIP 0.399893 LIN9 0.399956 BCAT1 0.400159 HERC4

Wang, et al. miR-202-3p targets ARL5A in CRC

0.400498 NUB1 0.400851 LYPLA1 0.40086 PSAT1 0.401007 ISG15 0.401178 SLC2A5 0.401244 TXK 0.401932 KIAA1267 0.402059 FAM12A 0.402315 LOC100132798 0.402474 CPEB1 0.402495 MRS2 0.402523 BCL7A 0.402529 FCER1A 0.402578 TAF12 0.403391 ODF3B 0.403817 IRX3 0.403927 ZFP91 0.404004 ZDHHC3 0.404068 KRTCAP2 0.40419 CLDN12 0.4044 TNIK 0.404446 UBA7 0.404991 ADAR 0.405012 ZBTB7C 0.406392 BAMBI 0.40643 TSEN54 0.406487 RPP30 0.407719 VPS53 0.408049 GRHL1 0.408203 DCLK1 0.408399 C4A /// C4B 0.408422 LOC728153 0.408661 RP11-257K9.7 0.408664 MATN2 0.408999 TTLL5 0.409076 BTN3A2 /// BTN3A3 0.409438 MTSS1L 0.409658 CHIC1 0.409965 RBBP9 0.410507 TBX6 0.410586 RRAD 0.410846 ANKLE1 0.411189 L1CAM

Wang, et al. miR-202-3p targets ARL5A in CRC

0.411502 PPP2R5C 0.411633 GCNT1 0.411965 C1orf201 0.412097 KIAA1467 0.412339 SYCE1 0.412403 PHF21A 0.412469 LOC100131914 0.412976 SLC2A3 0.413111 ARRDC3 0.413663 NOV 0.414626 TMEM135 0.414748 MGA 0.41501 ZNF587 0.416992 SLC30A1 0.417246 GATA3 0.41738 PPM1A 0.417626 HNF4G 0.41788 GBP3 0.418139 HECTD2 0.418181 AHNAK 0.418615 NACA2 0.418815 THTPA 0.418823 ZNF596 0.419166 ANUBL1 0.421109 LOC339290 0.421191 PTPN2 0.421232 PELO 0.421311 CLIC2 0.42175 NT5E 0.422038 ARMCX4 0.422345 FLJ20674 0.422458 AFAP1 0.423492 AVPI1 0.4236 ZADH2 0.423869 LOC652900 /// SEZ6L2 0.423872 ZNF28 0.423969 MKX 0.424208 C10orf47 0.424547 UCP3 0.424912 VAPB 0.425411 C11orf31 0.425857 GNG2 0.425931 LOC100132707

Wang, et al. miR-202-3p targets ARL5A in CRC

0.426239 RBM18 0.4266 ZNF236 0.42717 ACAA1 0.427426 FBXO4 0.42768 C9orf93 0.427707 METTL6 0.428463 S100P 0.428617 LOC100129502 0.428728 UBE2D3 0.428759 H2BFM /// H2BFXP 0.4291 CTDSP2 0.429801 MCTP2 0.430031 LOC401577 0.430158 C10orf119 0.430858 LOC647500 0.431136 BID 0.431205 AQP3 0.43235 ELL3 0.432363 HBS1L 0.432532 EPOR 0.432741 SLC2A14 /// SLC2A3 0.432866 CASP7 0.432974 TSC22D2 0.433048 TMEM173 0.433635 ARMC10 0.4338 ZNF577 0.434059 DOCK3 0.434111 B4GALT5 0.43478 CSRNP2 0.435023 EFCAB5 0.435051 GALNT12 0.435632 PTPRB 0.436023 MPL 0.436024 GTPBP5 0.436231 ZNF554 0.436489 RIPK2 0.436549 ABCD3 0.437081 GPR115 0.437336 PCTP 0.438126 UNC93B1 0.438594 ERCC8 0.438822 FLJ34208 0.438891 RGS10

Wang, et al. miR-202-3p targets ARL5A in CRC

0.440089 TFAP2A 0.440434 CDR2L 0.440485 FUCA1 0.440596 IFITM2 0.441267 TMED4 0.441872 CREB3L1 0.442062 MXRA8 0.442211 LOC100130886 /// TMEM164 0.443423 ZDHHC21 0.443492 LOC646014 0.444147 CXCL2 0.444883 LOC387895 0.445802 CRABP1 0.445842 ALDH5A1 0.446712 ETS2 0.447111 BIVM 0.447192 FAM86C 0.447349 XRCC4 0.447437 CDH8 0.447589 SBF2 0.448811 ZNFX1 0.44966 EMILIN2 0.449927 TSC22D3 0.450564 CXADRP1 0.45108 TRIM34 /// TRIM6-TRIM34 0.451389 UHRF1BP1 0.451745 FAS 0.451775 ZNF611 0.452877 C11orf1 0.453602 PPM1K 0.454128 SLC6A8 0.45419 LNPEP 0.454793 POU6F1 0.454824 AGK 0.455187 N4BP1 0.456431 KRT86 /// LOC100134394 0.457045 KIAA1609 0.457171 RAG1 0.457684 B2M 0.458212 ZNF396 0.458223 C14orf145 0.458432 FBF1 0.458797 LRRC7

Wang, et al. miR-202-3p targets ARL5A in CRC

0.459032 ALG12 0.459064 ATP10A 0.459452 EBF1 0.459656 LOC100134361 0.460148 RHEBL1 0.460304 G0S2 0.46085 APOL2 0.461835 CNOT6 0.46186 PLEKHA1 0.46214 C22orf36 0.462886 TET2 0.463761 INO80B 0.464108 LOC652147 0.464167 TRIM2 0.464195 LASS6 0.464375 MTHFSD 0.464674 ANXA8 /// ANXA8L1 /// ANXA8L2 0.464816 ZNF709 0.465053 LPCAT4 0.465226 TTC9 0.465503 LMLN 0.465963 MCC 0.46618 CACNB3 0.466693 ZNF296 0.46676 NPR1 0.467687 NFYA 0.468079 VEGFC 0.468198 BTN2A2 0.468422 HISPPD2A 0.468497 C12orf51 0.469654 MTF1 0.469683 SKIL 0.469711 TXNIP 0.469922 FOXO4 0.470033 ADAM19 0.47007 SPHK1 0.470359 STT3B 0.471122 LOC731484 0.471144 LOC148987 0.471264 BAIAP2L2 0.47132 ETNK2 0.471436 SPTLC3 0.471746 CSGALNACT1

Wang, et al. miR-202-3p targets ARL5A in CRC

0.471872 BACE2 0.471942 LOC90784 0.472003 NUPL1 0.472011 ZNF703 0.472193 TRIM62 0.472298 SUFU 0.472412 STK4 0.47248 DDX56 0.472861 LRIG3 0.473169 IVNS1ABP 0.473296 RIMKLB 0.473527 ZNF37B 0.473586 TRAF1 0.47445 CTSL1 0.474792 HCCS 0.474802 PER1 0.474862 TMEM145 0.475301 CALCB 0.475455 C6orf170 0.475476 APOL3 0.47585 C20orf70 0.475935 ZNF320 0.476414 SENP7 0.4765 GABARAPL1 /// GABARAPL3 0.476554 SCNN1G 0.476957 ZBTB4 0.477228 CA12 0.477406 IL24 0.477663 ZNF460 0.477665 SLC10A7 0.477953 CABC1 0.478337 POLR2C 0.478471 MVP 0.479158 ZNF788 0.479289 EPS15 0.479305 CARD16 /// CASP1 0.479548 BMP2K 0.479664 GANC 0.479871 HUNK 0.480046 ZNF37A 0.480192 CARD11 0.480749 PIK3C2B 0.480764 ZBED5

Wang, et al. miR-202-3p targets ARL5A in CRC

0.480807 PPIC 0.481123 CDC42SE1 0.481326 PRKY 0.481963 FAM63B 0.482176 UHRF1BP1L 0.482254 TPD52L1 0.482564 HSPG2 0.483448 VPS13D 0.484108 NCBP1 0.484184 ZNF673 0.484331 SLC16A1 0.484987 CTDSPL 0.485011 RPL5 /// RPL5P1 /// RPL5P34 /// SNORA66 0.48508 EFEMP1 0.485515 TRAF4 0.487255 TRIM25 0.487305 CD83 0.487437 SPIB 0.487451 KDM6A 0.487609 HIST1H4B 0.487651 SLC5A7 0.487736 TMEM67 0.487871 PARP10 0.487889 C1orf110 0.488284 INTS1 0.4883 FLJ39739 0.488711 TBC1D12 0.488718 C11orf17 /// NUAK2 0.48906 DNAL1 0.489295 DYRK2 0.489598 BCL2A1 0.489605 PNKD 0.489617 SCARF1 0.489776 PLXNC1 0.48998 TP53RK 0.490313 DNM3 0.490387 MMAA 0.490498 TNFRSF10C 0.490542 C1orf163 0.490771 NEDD9 0.491034 THSD1 /// THSD1P 0.491052 OCA2 0.491293 SGPP1

Wang, et al. miR-202-3p targets ARL5A in CRC

0.492297 DNAJC4 0.492362 FKSG83 0.492587 ZNF704 0.492858 C16orf87 0.493396 FBXW7 0.49343 FBLN1 0.494005 TSPAN2 0.494489 C1orf217 0.494533 CACNB2 0.494544 CA2 0.494597 CDKN1A 0.495211 KLHL28 0.495504 PSEN2 0.495609 ZNF607 0.496092 OLAH 0.496466 SEC23IP 0.496966 C3orf31 0.497177 FRMD3 0.497252 CEBPD 0.497973 SLC1A3 0.497984 LRRC58 0.498121 NCOA7 0.498271 WIPF1 0.498305 C19orf55 0.498506 ERAP2 0.499206 LOC100132116 0.499328 PVRL1 0.49937 ANGEL2 0.499416 PAPOLG 0.499492 GAS6 /// LOC100133684 0.499543 RIOK2 0.499748 CBR4 0.499907 XIAP 0.49996 KIAA1530

Wang, et al. miR-202-3p targets ARL5A in CRC

Supplementary Figures Figure S1.

A. The influence of miR-202-3p T:N expression ratios levels on the overall survival of CRC patients. A low T:N ratio indicated poor prognosis. B. The expression of miR-202-3p was examined in 6 different CRC cell lines. C. Upregulating or downregulating miR-202-5p expression in HCT-116 and LoVo cells did not affect their rates of proliferation. The cell numbers were determined using the CCK-8 assay after transfection. D. Transwell migration assays were performed using HCT-116 and LoVo cells infected with lenti-miR-202 or lenti-control viruses; miR-202 did not affect the migration of HCT-116 or LoVo cells.

Wang, et al. miR-202-3p targets ARL5A in CRC

Figure S2.

A. The effect of miR-202-3p on tumor formation in a nude mouse xenograft model. HCT-116 cells (1×106), transfected with miR-202-3p mimics or NC for 24 hours, were injected s.c. into the right flank of each nude mouse. The tumor formation rates were 8/8 (NT group) and 8/8 (miR-202-3p group). The tumor volume and weight of the miR-202-3p group was significantly decreased compared with the control group ( *: P<0.05, **: P<0.01, ***: P<0.0001). B, C. miR-202-3p expression was upregulated in post infected and transfected cells as well as in xenograft tumors compared with control. And ARL5A protein levels were isochronously decreased.

Wang, et al. miR-202-3p targets ARL5A in CRC

Figure S3.

A. The expression levels of ARL5A mRNA in HCT-116 and LoVo cells infected with miR-202-3p or control lentiviruses were determined by qRT-PCR. Beta-actin mRNA served as an internal control. B. The mRNA expression levels of target genes after silenced by specific si-RNAs were determined by qRT-PCR. Beta-actin mRNA served as an internal control. C. ARL5A and LRIG3 mRNA expression in 30 paired CRC and NCT samples. ARL5A mRNA was significantly upregulated in CRCs.

Wang, et al. miR-202-3p targets ARL5A in CRC

Figure S4.

A. Immunohistochemical staining of ARL5A in the CRC and NCT samples is shown. ARL5A protein is mainly localised in the cytoplasm. The scores (0, 1, 2 and 3) are based on the intensity of staining. B. ARL5A mRNA expression negatively correlated with miR-202-3p expression in the CRC tissues. C. Overall survival analysis was performed based on the expression level of ARL5A. The groups (T>N, T=N, T