Primary Hepatocyte Isolation and Culture

Human primary hepatocytes were obtained from the Liver Tissue Cell Distribution System at the University of Pittsburgh (Pittsburgh, Pennsylvania, USA). Primary mouse hepatocytes were isolated from 10-week-old WT C57BL/6J mice, as described previously (32 (link), 81 (link)). All cells were cultured in DMEM containing 10% FBS and then transfected with siRNAs described in the figure legends. The cells were then cultured in serum-free medium until they were harvested into RIPA buffer (Thermo Fisher Scientific, catalog 89900) with Halt Protease and Phosphatase Inhibitor Cocktail (Thermo Fisher Scientific, catalog 78444) for immunoblotting or into RNA lysis buffer (QIAGEN, catalog 79216) for mRNA quantification. Culture media were collected, snap-frozen in liquid nitrogen, and stored at –80°C until processing.