Protein Expression Analysis in Rat Liver

Protein extracts were obtained from liver samples of Ctrl, Fructose, HF, HF-F, and HF-F-D rats using standard procedure [16 (link)]. Aliquots of 50 μg of total protein from each sample were loaded into 4–15% pre-cast polyacrylamide gels (Bio-Rad, Milan, Italy) for Western blot analysis. Anti-PrxIII (LF-PA0030,Ab FRONTIER, Seoul, Korea), anti-PGC-1α (NBBP1-04676, Ab NOVUS, Centennial, CO, USA), anti-TFAM (74955), anti-COX-IV (48445), anti-Beclin-1 (37385), anti-LC3 (127414), and anti-β-actin (4970) (Cell Signaling, Danvers, MA, USA) were used as primary antibodies. The protein signals were detected by chemiluminescence (Clarity Western ECL substrate, Bio-Rad, Milan, Italy); the densitometric analysis was obtained using the Molecular Imager ChemidocTM (Bio-Rad, Milan, Italy), and data were normalized against β-actin expression.