Protein Isolation and Western Blot Analysis

Total protein was isolated from cells or tissues using RIPA buffer. Protein separation and western blot analysis were conducted as described earlier^{66 (link)}. GADD45α antibody (GTX54090, 1:1000) was from GeneTex. UCP1 (ab10983, 1:2000) and Perilipin (ab61682, 1:2,000) were from Abcam. FABP4 (E71703-98, 1:2000), GAPDH (EM1101, 1:5000) was from HuaBio. PPARγ (C26H12, 1:1000) was from Cell Signaling Technology (CST). Cocktail (45-8099, 1:2000) is from Thermo Fisher Scientific. PGC1α (sc-13067) was from Santa Cruz Biotechnology (Santa Cruz). The horseradish peroxidase (HRP)-conjugated secondary antibody (anti-rabbit IgG, 111-035-003 or anti-mouse IgG; 115-035-003, Jackson ImmunoResearch) was diluted 1:10,000. Immunodetection was performed using enhanced chemiluminescence western blotting substrate (Google Biotechnology, Wuhan, Hubei, China) and detected by ChemiScope3500 Mini System.