Sk hep 1

The HCC cell lines SK-HEP-1 and HEP3B were purchased from Procell (Wuhan, China). All cells were cultured in DMEM (Gibco) supplemented with 10% fetal bovine serum (Gibco), 1% streptomycin, and penicillin. Cells were transfected with lentivirus or plasmid purchased from Shanghai Genomeditech (Shanghai, China) and verified by DNA sequencing. Lentiviruses containing shNC (negative control, NC) and shSGOL2 were constructed using the vector pGMLV-SC5. The shRNA sequence used to target SGOL2 was as follows: 5′-GGTCAGAATTCCCTAACTTGT-3′. The pGMLV-SGOL2 plasmid contained the SGOL2 coding sequence, and the pGMLV-MAD2 plasmid contained the MAD2 coding sequence. For plasmid transfection, SK-HEP-1 and HEP3B cells were seeded in 12-well plates and then transfected with plasmids (4 mg per well) using Lipofectamine 2000 Reagent (Invitrogen) according to the protocols. Cells were harvested for analysis after 48 h.

All HCC cell lines were authenticated by STR profiles. HepG2 and SK-Hep-1 cell lines were authenticated by Procell Life Science & Technology Co, Ltd (Wuhan, China). Genomic DNA was extracted from HCC cells (1 × 10⁶) using PureLink Genomic DNA Mini Kit (Life K182001, USA), which was further amplified using PowerPlex^®18D System (Promega DC1802, USA), and sequenced by ABI3500 Genetic Analyzer (Life3500, USA). Hep3B cell lines were authenticated by BeNa Culture Collection (Beijing, China). Amplified genomic DNA was sequenced by ABI3730 XL Genetic Analyzer.

The human hepatoma cell lines, HepG2, Hep3B and SK-Hep-1 were purchased from Procell (Wuhan, China). The mice hepatoma cell lines, H22 and Hepa1–6 were purchased from the China Center for Type Culture Collection (Wuhan, China). All the cell lines were maintained in Dulbecco’s Modified Eagle Medium (DMEM)-high glucose supplemented with 12% FBS in the presence of 1% penicillin/streptomycin at 37 °C under the atmosphere of 95% air and 5% CO₂. The cell lines were identified by STR profiling and relative information is available in supplementary files.

For cell culture, HEK-293T and Jurkat cell lines was obtained from Haixing Biosciences (Suzhou, Jiangsu, China). The hepatocellular carcinoma cell lines (HepG2 and SK-Hep1) were obtained from Procell Life Science & Technology (Wuhan, Hubei, China). HEK-293T and SK-Hep1 cells were cultured in Dulbecco's Modified Eagle Medium (DMEM, Meilunbio, China) with 10% Fetal Bovine Serum (FBS, Standard Quality, OriCell, China). Jurkat cells were cultured in Roswell Park Memorial Institute-1640 (RPMI-1640, Meilunbio, China) with 10% Fetal Bovine Serum (FBS, Standard Quality, OriCell, China). HepG2 cells were cultured in Minimum Essential Medium (MEM, Meilunbio, China) with 10% Fetal Bovine Serum (Standard Quality, OriCell, China). All cells were grown at 37 °C with 5% CO₂. For transfection, cells were transiently transfected with plasmid, siRNA, and shRNA using Lipo6000 Transfection Reagent (Beyotime, Shanghai, China) according to the manufacturer's protocol. For lentivirus infection, lentivirus containing pLVX-BCLAF1-Puro, pLVX-shBCLAF1-Puro, pLVX-SPOP-Puro, pLVX-shSPOP-Puro, and corresponding Control group lentivirus were purchased from Obio Technology (Shanghai, China) and were used to infect HepG2 and SK-Hep1 cells according to the manufacturer's protocol. Infected cells were then subjected to puromycin selection (5 μg/mL), and stable transfection of cells was confirmed by western blot.

The human liver cancer cells SK-HEP-1 was obtained from the Procell Life Science & Technology Co., Ltd (Wuhan, China) and was authenticated through STR genotyping. The cell lines were tested periodically for mycoplasma contamination during the experiment and were confirmed negative. Cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, CA, USA) and 1% penicillin–streptomycin solution (Beyotime, Shanghai, China).
4MOD was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). In animal experiments, 0.25% polyoxyl castor oil (Yuanye, Shanghai, China) and 0.25% EtOH were used to dissolve 4MOD. A rabbit polyclonal antibody anti-GADD45G was obtained from Bioss (Beijing, China).

Human hepatocytes (WRL68 cells) and HCC cells (HepG2, SK-Hep-1, Huh7, and SMMC772) were obtained from the American Type Culture Collection (Manassas, VA, USA). WRL68, SK-Hep-1, and HepG2 cells were cultured in minimum essential medium (Procell, Wuhan, China, PM150410) supplemented with 10% fetal bovine serum (FBS, HyClone, USA, SH30396), while SMMC7721 and Huh7 cells were, respectively, cultured in RPMI-1640 medium and DMEM (Procell, PM150110B) supplemented with 10% FBS.