Western Blot Analysis of PPARG

Western blot analysis was implemented as described previously (Ma et al., 2019 (link)). In brief, aliquots of total protein extract (20 μg) from cells were loaded and resolved by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After electrophoresis, proteins were transferred onto polyvinylidene difluoride membranes (PVDF; EMD Millipore, Billerica, MA, United States). Blots were blocked with 5% non-fat milk in PBST (PBS containing 0.1% TWEEN-20) for 1 h at room temperature, and the membrane was incubated with primary antibody against PPARG (Abcam, ab178860) at 4°C overnight. The membrane was then washed three times with PBST and exposed to horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (1:2000 dilution, Cell Signaling Technology, United States) for 1 h at room temperature. After three washes with PBST, antibody binding was detected with enhanced chemiluminescence substrates (Millipore, United States) and visualized with an ImageQuant LAS 4000 mini-densitometer (GE Healthcare Life Sciences, United States).