Adenosine triphosphate (atp)

Male C57BL/6 mice (6-8 weeks) were provided by the Laboratory Animal Center of Chongqing Medical University. All of the animals were placed in a specific pathogen-free environment and all surgical procedures and care administered to the animals were approved by the institutional ethic committee. The mice received the intraperitoneal injection of a mixture of carbon tetrachloride (CCl4, 50%) and oil (50%) at a dose of 2 ml/kg body weight. They were scarified after the CCl4 injection for 24 h. 0.2 ml G-Rg1 (4 mg/ml, Jicui Biotechnology Co., Ltd., Yunnan, China) was given after the CCl4 injection.
The suppression of autophagy was achieved by the tail vein injection of 3-methyladenine (3-MA, 1 mg/kg, Sigma) 2 h before CCl4 exposure, while intraperitoneal injection of autophagy agonist rapamycin (RPA, 1 mg/kg, Sigma) was given 0.5 h before CCl4 exposure. Intraperitoneal injections of MCC950 (10 mg/kg, MedChem Express, Shanghai, China) or 0.3 ml ATP (100 Mm, Santa Cruz Biotechnology) was used to inhibit or activate NLRP3 inflammasome.

Ectonucleotidase activity was measured in the cells as described previously [32 (link)]. Briefly, cells were seeded at 10,000 cells/cm² in 24-well plates and then incubated for 24 h. ATP, ADP and AMP (1 mM; Santa Cruz) were then added in phosphate-free buffer and incubated for either 1 h (ATP and ADP) or 30 min (AMP). Supernatant was harvested for protein quantification (BCA assay; Thermo Fisher, Waltham, MA, USA), inorganic phosphate quantification (malachite green assay kit, according to the manufacturer’s instructions; Sigma Aldrich) or adenosine detection.