Western Blot Analysis of Liver Proteins

Protein extraction was performed in liver for Western Blot analysis, as previously described [26 (link),27 (link)]. About 30 μg of proteins were loaded and specific antibodies were used to evaluate PPARα (ab61182), SREBP-1 (ab28481) (1:250 and 1:1000 dilution respectively; Abcam, Cambridge, MA, USA), p-AMPK-α (2535), and β-actin (4970) (1:1000 dilution each; Cell Signaling, Beverly, MA, USA) which was used as loading control. The images were obtained using specific software (C-DiGit Blot Scanner with Image Studio 4.0 software, LI-Cor, Lincoln, NE, USA), and densitometric data were expressed as integrated intensity, following evaluation of a region of interest (ROI) identical for all samples within the same blot.