Western blotting
T-PER (Thermo Fisher Scientific Inc.) and Halt protease and phosphatase inhibitor cocktail (Thermo Fisher Scientific Inc.) were mixed at a ratio of 99:1 to prepare the lysis buffer. Liver samples were homogenized in the lysis buffer and centrifuged at 10,000 × g for 10 min. The supernatants were collected, each sample was mixed with Lane marker reducing sample buffer (Thermo Fisher Scientific Inc.) at a ratio of 4:1, and heated at 95 ºC for 5 min to prepare the blotting samples. The prepared samples were separated using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Proteins were transferred from the gel onto a PVDF membrane using an iBlot Dry Blotting System (Thermo Fisher Scientific Inc.). The primary antibodies reactions [anti-Vinculin antibody (Cat# 13901S, RRID:AB_2728768) was purchased from Cell Signaling Tech, Inc.; anti- NQO-1 antibody (Cat# ab34173, RRID:AB_2251526), anti-Glutamate-cysteine ligase catalytic subunit (GCLC) antibody (Cat# ab207777, RRID: none yet assigned), and anti-alpha smooth muscle actin (αSMA) antibody (Cat# ab32575, RRID:AB_722538) were purchased from Abcam Inc. (Cambridge, UK) ] were performed using the iBindTM Western Device (Thermo Fisher Scientific Inc.). The bound primary antibodies were detected using an HRP-conjugated goat anti-rabbit IgG secondary antibody (Cat # HAF008, RRID: AB_357235; R&D Systems Inc., Minneapolis, MN). Then, AmershanTM ECL Prime western blotting detection reagent (GE Healthcare Inc., Chicago, IL) and ChemiDocTM XRS Plus (Bio-Rad Laboratories Inc., Hercules, CA) were used for protein detection. All immunological procedures were performed according to the recommendations of theBritish Journal of Pharmacology (Alexander et al., 2018).