Gssg assay kit

Specific pathogen-free male C57Bl/6J mice (weight 18–22 g) were supplied from the Shanghai Laboratory Animal Center, Chinese Academy of Sciences. The animals were housed. All experiments abided by institutional ethical guidelines of the Animal Care and Use Committee (Shanghai Institute of Materia Medica, Chinese Academy of Sciences, China). For drug administration, acetaminophen was dissolved in normal saline, whereas dabrafenib and vemurafenib were dissolved in pH 8.0 distilled water containing 0.5% HPMC and 0.2% Tween 80. Overnight-fasted mice were given dabrafenib (300 mg/kg or 100 mg/kg) or vemurafenib (300 mg/kg) by oral gavage 1 h before the intraperitoneal injection of 300 mg/kg acetaminophen. The mice were killed 6 h post the acetaminophen treatment. Plasma alanine aminotransferase and aspartate aminotransferase were measured with a Cobas C501 Chemistry Analyzer (Roche, Basel, Switzerland). Plasma IL-1β and TNF-α were measured by using the mouse IL-1β ELISA kit and the mouse TNF-α ELISA kit (MultiSciences, Bellingham, WA, USA), respectively. Liver GSSG was measured by using a GSSG Assay kit (Beyotime). To evaluate the changes in liver histology, sections of paraformaldehyde-fixed liver tissues were stained with hematoxylin and eosin and photographed under an optical microscope. The standard TUNEL staining was carried out to evaluate the DNA breaks in liver cells.

Glutathione (GSH)/glutathione disulfide (GSSG) Assay Kit (Beyotime Institute of Biotechnology, Jiangsu, China) was used to detect the content of GSH/GSSG in cells. The cell processing method was used as described above, and according to the instructions the absorbance at 412 nm was read with a microplate reader.