Western Blot Analysis of Adipose and Energy Metabolism

The standard procedure for Western blotting was performed using the following antibodies: anti-adipose triglyceride lipase antibody (abcam, ab109251, Cambridge, MA, USA), anti-AMPKα antibody (CST, mAb #2603, Danvers, MA, USA), anti-phospho-AMPKα antibody (CST, mAb #2535), anti-SREBP1 antibody (Novus Biologicals, NB-100-2215SS, Littleton, CO, USA), anti-β-actin antibody (Boster, BM0627, Wuhan, China). Western blot analysis was done as described elsewhere [43 (link)]. Briefly, whole-tissue and cell lysates were prepared, and protein concentrations were measured using the bicinchoninic acid (BCA) method. Proteins were separated on the Bio-Rad Mini-PROTEAN Tetra gel system (BioRad, Hercules, CA, USA). The proteins were transferred to PVDF membranes and probed. Equal loading of the protein was confirmed by β-actin.