Maintenance of Liver Cancer Cell Lines

Liver cancer cell lines Hep3B, HepG2, Huh7, PLC, SKHep1 and Snu387 were maintained as previously described^{44 (link)}. Briefly, Hep3B, HepG2, Huh7, PLC and SKHep1 lines were cultured in low glucose DMEM while Snu387 line was cultured in RPMI, and in both cases supplemented with 10% fetal bovine serum, 100 U/mL penicillin-streptomycin and 0.1 mM non-essential amino acids. In addition, Huh7 cells stably overexpressing GFP were supplemented with 200 μg/mL geneticin G-418 sulfate (Thermo Fisher Scientific, USA) and no additional antibiotics^{34 (link)}. Cells were grown at 37 °C in an incubator supplied with 5% CO₂. Cells reaching confluency (every 3 to 4 days) were split into new plates containing fresh media.