Protein Extraction and Western Blot Analysis

Total cell lysates were prepared using radioimmunoprecipitation assay buffer (RIPA) or 2x Laemmli buffer supplemented with proteinase inhibitor cocktail (Sigma, USA) and phosphatase inhibitor cocktail (Sigma,USA). Liver samples were lysed and sonicated in T-PER (Thermo Fisher Scientific, Rockford, USA) per manufacturer’s protocol. Protein concentrations were determined using abicinchoninic acid (BCA) protein assay kit (Thermo Fisher Scientific, Rockford, USA). Protein levels were analyzed by Western blot, as previously described^{31 (link),32 (link)}. β-Actin or GAPDH levels were also determined as controls. Protein bands were detected by chemiluminescence (Kodak Digital Science Image Station) or infrared fluorescence (Odyssey Image station and Image Studio, LI-COR Biosciences, Lincoln, NE, USA).