Hepg2 cells

The HepG2 cells were obtained from Boster Biological Technology Co., Ltd (Wuhan, China). In our study, hPXR-silenced HepG2 and hCAR-silenced HepG2 cell models were successfully constructed by respectively transfecting shPXR and shCAR plasmids using the SuperFectin II DNA transfection reagent (PUFEI Biology) following the manufacturer’s instructions. The HepG2 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS at 37°C in a humidified 5% CO₂ atmosphere. Cell passages were performed with 0.25% trypsin-EDTA (Gibco, Life Technologies) when cells reached 80–90% confluency. For the treatments, OA, UA, RIF, or CITCO was dissolved in dimethyl sulfoxide (DMSO) for the final stock solution concentrations of 20, 20, 50, and 50 mM, respectively and diluted with DMEM containing 10% FBS. The final concentration of DMSO never exceeded 0.1% (v/v) in the medium.

BDDE was kindly provided by Dr. Fan, Institute of Oceanology, Chinese Academy of Sciences. HepG2 cells were purchased from BOSTER, Ltd. (Wuhan, China). Dulbecco’s modified Eagle’s medium (DMEM) and fetal bovine serum (FBS) were purchased from Invitrogen (Carlsbad, CA, USA). MTT, metformin and insulin were purchased from Sigma-Aldrich (St. Louis, MO, USA). The antibody directed against total IRβ, phospho-IRβ, total IRS-1, phospho-IRS1/2, total PI3K, phospho-PI3K, total Akt, and phospho-Akt (Ser473) was purchased from Cell Signaling Technology (Danvers, MA, USA). The antibody directed against GAPDH was purchased from Abcam Trading Company Ltd (Shanghai, China). A glucose assay kit was purchased from Solarbio (Beijing, China). All the other reagents were purchased from Sigma, unless otherwise indicated.