Liver cancer HepG2 cells, purchased from the American Type Culture Collection, were cultured in high-glucose DMEM (GE Healthcare Life Sciences) with 10% fetal bovine serum (Clark Bioscience) and 1% penicillin/streptomycin at 37°C in an atmosphere containing 5% CO2. The cells were dissociated with 0.25% trypsin (w/v) and 0.52 mM EDTA [M&C Gene Technology (Bejing) Ltd.] and routinely sub-cultured at 80% confluency.
Cell cultures were treated with various concentrations of apigenin in DMSO as the carrier solvent. Palmitic acid binds to fatty-acid-free BSA (Beijing Solarbio Science & Technology Co., Ltd.). In brief, palmitic acid was dissolved in 1X PBS and a 250 mM stock solution was obtained following various cycles of incubation in a water bath at 70°C and vortexing. The stock solution was then added to serum-free DMEM containing 5% fatty-acid-free BSA to obtain a 250 µM palmitic acid solution, and the resulting diluted solution was used for the cell treatments (16 (link),17 (link)). AMPK inhibitor compound C was diluted with DMSO to a final concentration of 10 µM and was used to verify the signaling pathway.
Cell cultures were treated with various concentrations of apigenin in DMSO as the carrier solvent. Palmitic acid binds to fatty-acid-free BSA (Beijing Solarbio Science & Technology Co., Ltd.). In brief, palmitic acid was dissolved in 1X PBS and a 250 mM stock solution was obtained following various cycles of incubation in a water bath at 70°C and vortexing. The stock solution was then added to serum-free DMEM containing 5% fatty-acid-free BSA to obtain a 250 µM palmitic acid solution, and the resulting diluted solution was used for the cell treatments (16 (link),17 (link)). AMPK inhibitor compound C was diluted with DMSO to a final concentration of 10 µM and was used to verify the signaling pathway.
