Western Blot Analysis of Rg3 Signaling

Western blot analysis was performed as previously described [19 (link)]. Briefly, MPBAs were treated with the indicated doses of Rg3 for two days. Total proteins were extracted using lysis buffer containing protease and phosphatase inhibitors, followed by processing for SDS-PAGE. The blots were incubated with the following primary antibodies as appropriate: PPARγ, perilipin, ERO1L, Phospho-eIF2α, CHOP, Phospho-Akt, total Akt, cleaved caspase3, total caspase3, and β-actin (all antibodies purchased from Cell Signaling Technology). An ECL Prime Western Blotting System (GE Healthcare) and ImageQuant LAS4000 (GE Healthcare) were used to detect protein bands. Densitometry analysis was performed using Image J software (NIH, Bethesda, MD, USA).