Snu423

The SNU423, SNU449, and MHCC-97H cell lines were purchased from ATCC. The WT and TSC1^–/– MEF lines were gifts from X.F. Steven Zheng. SNU423 and SNU449 cells were cultured in RPMI 1640 medium (Gibco, USA), while MHCC-97H cells and MEFs were maintained in high-glucose Dulbecco’s modified Eagle’s medium (DMEM; Gibco). Fetal bovine serum (FBS; 10%, Gibco) and penicillin–streptomycin solution were added to the medium during cell culture. The cells were incubated at 37 °C in a humidified chamber containing 5% CO₂. Rapamycin, decitabine, and MG-132 were purchased from Selleck Chemicals (USA); CHX was purchased from Sigma (USA).

The human liver cancer cell lines (HepG2, Huh7, Hep3B, SNU-423, HCC-LM3, MHCC-97H, SK-Hep1, and SMMC-7721) and two human immortalized normal liver cell lines (LO2 and THLE-2) were obtained from the Institute of Biochemistry and Cell Biology at the Chinese Academy of Sciences (Shanghai, China) or American Type Culture Collection (ATCC) and supplemented with Dulbecco's modified Eagle's medium (DMEM; Gibco, USA) mixed with 10% fetal bovine serum (FBS; Gibco, USA) and 1% antibiotics (penicillin and streptomycin) at 37°C in a humidified incubator containing 5% CO₂. All the cell lines have been tested for contamination before cell experiments. The IκBα phosphorylation inhibitor Bay11-7082 was purchased from Selleck (S2913, Shanghai, China) and dissolved in dimethyl sulfoxide (DMSO; as a vehicle).