Gapdh

A detailed description of protein extraction and Western blotting procedures can be found in our previous report [5 (link)].
In brief, muscle samples were loaded and separated on a 10% polyacrylamide gel, followed by transfer to a nitrocellulose membrane (Santa Cruz Biotechnology, Inc., Sanford, ME, USA, #sc-3724), after which membranes were incubated in a blocking buffer (TBS-T: 4% non-fat milk powder; Tris-buffered saline, pH 7.4; and 0.1% Tween 20). The membranes were then incubated with primary and secondary antibodies and washed in TBS-T. The primary antibodies used were GAPDH (1:10,000, Applied Biological Materials Inc., Richmond, BC, Canada, # G041), Lamin B1 (1:500, Abcam, Cambridge, MA, USA, # ab16048), MEF2-D (1:1000, EMD Millipore, Temecula, CA, USA, # AB2263), acetyl-Histone H3 (1:1000, EMD Millipore, Temecula, CA, USA, # 06-599), total Histone H3 (1:1000, Cell Signaling Technology, Danvers, MA, USA, # 9715), HDAC4 (1:500, Cell Signaling, Danvers, MA, USA, #2072), HAT P300 (1:500, Abcam, Cambridge, MA, USA, # ab231010).
Secondary HRP-conjugated antibodies (1:30,000) to rabbit or mouse immunoglobulins were from Santa Cruz Biotechnology, CA, USA. Protein bands were detected and quantified using Clarity Western ECL Substrate (Bio-Rad Laboratories, Hercules, CA, USA, #170-5061) and C-DiGit Blot Scanner (LI-COR Biotechnology, Lincoln, NE, USA).

Western blotting was performed as described previously (Lu et al., 2017 (link)). In brief, fresh liver tissues were homogenized in RIPA and PMSF. 5 × SDS-PAGE loading buffer was added, followed by protein degeneration via boiling for 5 min. The loaded protein were separated by 10 or 12% SDS-PAGE, and transferred to nitrocellulose membrane (NC). The membranes were incubated for 1 h in non-fat dry milk in TBST. CYP1A1 (1:500 diluted, Sangon Biotech. Shanghai, China), CYP1A2 (1:400 diluted, Sangon Biotech, Shanghai, China), CYP3A4 (1:200 diluted, Sangon biotech. Shanghai, China), CYP2A6 (1:200 diluted, ImmunoWay Biotechnology Company, Suzhou, China), Nrf2 (1:500 diluted, Wanleibio Co., Ltd., Harbin, China), GSTA3, (1:200 diluted, Life Span Biosciences, Inc., Shanghai, China), GSTM2 (1:800 diluted, BBI Life Sciences, Canada), and β-actin (1:1000 diluted, Applied Biological Materials Inc., Canada) or GAPDH (1:100 diluted, Applied Biological Materials Inc., Canada) protein was detected using specific primary antibodies incubated overnight at 4°C. Secondary anti-mouse or anti-rabbit horseradish peroxidase conjugated IgG was used for 1 h, and immune-complexes were detected using enhanced chemiluminescence (ECL) detection. Signals were visualized on a computer using Image J software (National Institute of Health, United States).