“…The membrane was blocked at room temperature in 5% BSA for 2 h, and the membrane was incubated with primary antibodies (UCP1 primary antibody, rabbit 1:1000, Abcam, Cambridge, UK; PGC1α primary antibody, rabbit, 1:1000, Abcam, Cambridge, UK; ACTIN primary antibody, mouse, 1:1000, Cambridge, UK; TUBULIN primary antibody, rabbit, 1:1000, Cambridge, UK) overnight at 4 °C. The following day, the membrane was washed in TBST three times, 10 min each, and the membrane was incubated with secondary antibodies for 2 h. The protein bands were visualized by chemiluminescence (Bio-Rad, Hercules, CA, USA) [34]. …”