KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| EphA2 (C-3) | Santa Cruz | sc-398832 |
| EphA2 (phospho Y588) (D7X2L) | Cell signaling | 12677S; RRID:AB_2797989 |
| EphA2 (phospho S897) (D9A1) | Cell signaling | 6347S; RRID:AB_11220420 |
| Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody | Thermo Fisher Scientific | 31432; RRID:AB_228302 |
| Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody | Thermo Fisher Scientific | 31462; RRID:AB_228338 |
| Goat anti-Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Biotin | Thermo Fisher Scientific | 31822; RRID:AB_228337 |
| Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Biotin | Thermo Fisher Scientific | 31802; RRID:AB_228301 |
| AKT (phospho S473) (D9E) | Cell signaling | 4060S; RRID:AB_2315049 |
| AKT (phospho T308) (244F9) | Cell signaling | 4056S; RRID:AB_331163 |
| AKT | Cell signaling | 9272S; RRID:AB_329827 |
| β-Actin | Sigma-Aldrich | A5441; RRID:AB_476744 |
| GADPH (GA1R) | Thermo Fisher Scientific | MA5–15738; RRID:AB_10977387 |
| GP130 | R&D | MAB2281; RRID:AB_2296041 |
| STAT3 (phospho Y705) (D3A7) | Cell signaling | 9145S; RRID:AB_2491009 |
| STAT3 (124H6) | Cell signaling | 9139; RRID:AB_331757 |
| MET (D1C2) | Cell signaling | 8198; RRID:AB_10858224 |
| β-Catenin | BD Biosciences | 610153; RRID:AB_397554 |
| Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488 | Thermo Fisher Scientific | A-11001, RRID:AB_2534069 |
| Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor 594 | Thermo Fisher Scientific | A-11037; RRID:AB_2534095 |
| Alpha-1-Fetoprotein Pathology Antibody | Agilent | A0008; RRID:AB_2650473 |
| Ki-67 (D3B5) | Cell signaling | 12202; RRID:AB_2620142 |
| JAK1 (phospho Y1034/1035) (D7N4Z) | Cell signaling | 74129; RRID:AB_2799851 |
| Jak1 (6G4) | Cell signaling | 3344, RRID:AB_2265054 |
| Bacterial and virus strains | ||
| MAX Efficiency Stbl2 Competent Cells | Thermo Fisher Scientific | 10268019 |
| MAX Efficiency DH5α Competent Cells | Thermo Fisher Scientific | 18258012 |
| Biological samples | ||
| Normal + HCC human liver samples | Mayo Clinic Rochester; Dr. Lewis R. Roberts | https://www.cancer.gov/about-nci/organization/ccg/research/structural-genomics/tcga |
| Chemicals, peptides, and recombinant proteins | ||
| ALW-II-41–27 | APExBIO | A3165 |
| Critical commercial assays | ||
| AlamarBlue® | BIO-RAD | BUF012B |
| GeneJET Endo-Free Plasmid Maxiprep Kit | Thermo Fisher Scientific | K0861 |
| RNeasy Mini Kit | QIAGEN | 74106 |
| Duolink® In Situ Red Starter Kit Mouse/Rabbit | Millipore Sigma | DUO92101 |
| Deposited data | ||
| RNaseq EphA2 knockdown | This Paper | Database: GEO, GSE141880 |
| RNaseq expression analysis of HCC and normal liver tissue | Database: TCGA | https://www.cancer.gov/about-nci/organization/ccg/research/structural-genomics/tcga |
| Experimental models: cell lines | ||
| Huh7 | JCRB | JCRB0403 |
| Hep3B | ATCC | HB-8064 |
| PLC/PRF/5 | ATCC | CRL-8024 |
| SNU387 | ATCC | CRL-2237 |
| SNU423 | ATCC | CRL-2238 |
| SNU449 | ATCC | CRL-2234 |
| SNU475 | ATCC | CRL-2236 |
| Experimental models: organisms/strains | ||
| NSG-A2 | Jackson Laboratory | 009617 |
| C57BL/6 | Jackson Laboratory | 000664 |
| Oligonucleotides | ||
| Primer sequences for qRT-PCR | Table S3B | N/A |
| Human EphA2 shRNAshEphA2–1: CCGGCCATCAAGATGCAGCAGTATACTCGAGTATACTGCTGCATCTTGATGGTTTTTG | Millipore Sigma | TRCN0000231647 |
| Human EphA2 shRNAshEphA2–2: CCGGGATAAGTTTCTATTCTGTCAGCTCGAGCTGACAGAATAGAAACTTATCTTTTTTG | Millipore Sigma | TRCN0000195734 |
| Human EphA2 shRNAshEphA2–5: CCGGTCGGACAGACATATAGGATATCTCGAGATATCCTATATGTCTGTCCGATTTTTG | Millipore Sigma | TRCN0000231648 |
| Human JAK1: siGENOME SMARTpool siRNA | Dharmacon/Horizon | SO-2796483G |
| Human JAK1: CCACAUAGCUGAUCUGAAA | Dharmacon/Horizon | D-003145–05 |
| Human JAK1: UGAAAUCACUCACAUUGUA | Dharmacon/Horizon | D-003145–06 |
| Human JAK1: UAAGGAACCUCUAUCAUGA | Dharmacon/Horizon | D-003145–07 |
| Human JAK1: GCAGGUGGCUGUUAAAUCU | Dharmacon/Horizon | D-003145–08 |
| Mouse EphA2 sgRNAsgEphA2–1: CAACGTGGTATCCGGCGACC | Dr. Feng Zhang; Cong et al., 2013 | MGLibA_16318 |
| Mouse EphA2 sgRNAsgEphA2–2: TTCGCTGTCGAAGCACGCAA | Dr. Feng Zhang; Cong et al., 2013 | MGLibA_16319 |
| Mouse EphA2 sgRNAsgEphA2–3: CTGCTGACCGTGATCTCGTC | Dr. Feng Zhang; Cong et al., 2013 | MGLibA_16320 |
| Recombinant DNA | ||
| pCMV-dR8.2 dvpr | Stewart et al., 2003 | 8455; RRID:Addgene_8455 |
| pCMV-VSV-G | Stewart et al., 2003 | 8454; RRID:Addgene_8454 |
| pLKO.1-puro-shEphA2–1 | Millipore Sigma | TRCN0000231647 |
| pLKO.1-puro-shEphA2–2 | Millipore Sigma | TRCN0000195734 |
| pLKO.1-puro-shEphA2–5 | Millipore Sigma | TRCN0000231648 |
| FG12 | Qin et al., 2003 | 14884; RRID:Addgene_14884 |
| FG12-CMV-CA-AKT | Dr. Mitchell Denning, LUC | Yadav and Denning, 2011 |
| pMD2.G | Dr. Didier Trono | 12259; RRID:Addgene_12259 |
| pLX304 | Yang et al., 2011 | RRID:Addgene_25890 |
| pEXP304-STAT3/V5 | Sinnberg et al., 2016; DNASU | HsCD00443857; PMID: 27428425 |
| pEXP304-FLAG/V5 | Dr. Takashi Shimamura, UIC | N/A |
| pT3-CAT | Khanna et al., 2004; Shang et al., 2015 | N/A |
| pT3-MET | Khanna et al., 2004; Shang et al., 2015 | N/A |
| pT3-GFP | Shang et al., 2015 | N/A |
| pT3-EphA2 | This Paper | N/A |
| pX330-U6-Chimeric_BB-CBh-hSpCas9 (PX330) | Dr. Feng Zhang; Cong et al., 2013 | Addgene plasmid # 42230; RRID:Addgene_42230 |
| pX330-sgEphA2–1 | This Paper | N/A |
| pX330-sgEphA2–2 | This Paper | N/A |
| pX330-sgEphA2–3 | This Paper | N/A |
| Software and algorithms | ||
| PRISM | GraphPad Software | Version 7 |
| GEPIA | Tang et al., 2017 | http://gepia.cancer-pku.cn/ |
| ImageJ | NIH | https://imagej.nih.gov/ij/ |
| GSEA | Broad Institute | http://www.gsea-msigdb.org/gsea/index.jsp |
| Bitplane Imaris | Oxford Instruments | https://imaris.oxinst.com/packages |