Quantitative Protein Analysis of Liver and Adipose

Protein was extracted from frozen liver and epididymal white adipose tissues (eWAT). Nuclear protein was prepared from mouse livers as described previously (Alam et al., 2017 (link)). Protein concentration was determined using a Pierce BCA Protein Assay kit (Thermo Fisher Scientific, MA). Western blotting was performed as described previously (Liu et al., 2020 (link)). Primary antibodies (1:500–1:1,000) against Phospho-HSL, ATGL, PPAR-γ, CPT-1a, Nrf2, BAX, BCL-2, Adiponectin, Cleaved-Caspase3, pro-Caspase3 and CYP7A1 were from Cell Signaling Technology (MA, United States). β-Actin and Lamin B antibody were the reference of total protein and nuclear protein, respectively, and purchased from Abcam (Cambridge, MA, United States). Densitometric analysis was performed using UN-SAN-IT Gel (Silk Scientific, Orem, UT) software.