Hcclm3

The four HCC cell lines, Huh-7 (AW-CCH089), SK-Hep-1 (AW-CCH036), HCC-LM3 (AW-CCH036), and HepG2 (AW-CCH024), were all purchased from Abiowell. Huh-7 cells and HCC-LM3 cells were cultivated in Dulbecco’s modified Eagle medium (DMEM, D5796-500ML, Sigma, USA), and SK-Hep-1 and HepG2cells were cultivated in a minimal essential medium (MEM, 11,095,080, Gibco, USA). These media were supplemented with 10% fetal bovine serum (FBS, 10,099,141, Gibco, USA) and 1% Penicillin/Streptomycin (AWI0070a, Abiowell, China). Cells were cultured in a saturated humidity incubator (DH-160I, SANTN, China) with 5% CO₂ at 37℃. The sh-METTL14, oe-METTL14, sh-CHOP, and oe-CHOP plasmids, along with their respective control plasmids, were procured from HonorGene. They were transfected into SK-Hep-1 and HCC-LM3 cells by using Lipofectamine 2000 (11,668,019, Invitrogen, USA).

All the cells were purchased from China Center for Type Culture Collection (CCTCC). Human hepatocellular carcinoma cells (SMMC-7721, SK-hep-1, HCC-LM3, Huh-7) and human immortalized normal hepatocytes (L-02) 19 were maintained in DMEM (Sigma, USA), MEM (Gibco, USA), or RPMI 1640 (Sigma, USA) containing 10% fetal calf serum (Gibco, USA) at 37 °C in a humidified incubator (Thermo Scientific, USA) with 5% CO2.

HL7702, Hep3B, MHCC97L and MHCC97H cells were obtained from the Cobioer Bioscience Company (Nanjing, China), and HCCLM3 cells were obtained from China Center for Type Culture Collection (Wuhan, China). HL7702 cells were cultured in 1640 medium (Life Technology, Carlsbad, CA, USA), and Hep3B, MHCC97L, MHCC97H and HCCLM3 cells were cultured in high‐glucose DMEM (Life Technologies Corporation, Carlsbad, CA, USA) and supplemented with 10% FBS (Life Technologies Corporation) in a humidified atmosphere of 5% CO₂. For GSK3β inhibition, MHCC97L and HCCLM3 cells were firstly incubated with 1 or 4 μg/ml BS‐I (Sigma‐Aldrich, St Louis, MO, USA) for 6 hrs, and then, 0.2 μM CHIR99021 (Selleckchem, Houston, TX, USA) or 4 mM LiCl (Sigma‐Aldrich) was added to the medium.

HL7701, HL-7702, HepG2, Huh7, Sk-Hep1, Bel7402, HLE, HLF and Alex cell lines were purchased from China Center for Type Culture Collection (CCTCC, Wuhan, China). MHCC-97 H and HCC-LM3 cell lines were obtained from Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China. 293T cells were purchased from the American Type Culture Collection. Cells were cultured in DMEM (Hyclone, Logan, UT, USA) supplemented with 4.5 g/L glucose and 10% FBS (Gibco; Thermo Fisher Scientific, Inc.). All cell lines have been tested for their authenticity. Transfection were performed using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacture’s instruction. For lentivirus production, pLKO.1 vector and pLKO.1-shSHC4 or pLenti-CMV-GFP vector and pLenti-CMV-GFP-SHC4 (6 µg), psPAX2 (4.5 µg), pMD2.G (1.5 µg) were purchased from Addgene, Inc. (Cambridge, MA, USA) and were co-transfected into 293T cells. The virus-containing supernatants were collected and filtered 48 h after transfection. Freshly made virus supernatants supplemented with 8 µg/mL polybrene (Sigma-Aldrich) were added to exponentially growing HepG2, Huh7 or HCC-LM3 cells. After 8 h, fresh medium was added. SHC4 stable overexpression or knockdown cells were achieved by 1-week puromycin (5 µg/mL, Ann Arbor, MI, USA) selection.