Mcc950 cp 456773

Antibodies used targeted mouse IL-1β (AF-401-NA, R&D Systems), mouse gasdermin D (ab209845, Abcam), ASC (AL177, Adipogen), mouse IL-1α (AF-400-NA, R&D Systems), human calpain 1 (ab39170, Abcam), human calpain 2 (ab39165, Abcam), and β-actin-HRP (A3854, Sigma). Pharmacological agents used were punicalagin (Sigma), glycine (Sigma), NBC6 (synthesized in house (17 (link))), MCC950 (CP-456773, Sigma), Z-VAD-fluoromethyl ketone (Merck), calpain inhibitor III (Merck), nigericin (Sigma), adenosine triphosphate (Sigma), ionomycin (Sigma), monosodium urate crystals (InVivoGen), calcium pyrophosphate dihydrate crystals (InVivoGen), and IL-Ra (Kineret®, Amgen). All other materials were from Sigma-Aldrich unless specified.

Different reagents and their sources used in this study were: ATP, nigericin, BSA fatty acid-free and low endotoxin, palmitic acid, doxycycline, MCC950 (CP-456773), and the authentic standards citric acid, malic acid, fumaric acid, succinic acid, ketoglutaric acid, pyruvic acid and lactic acid were all purchased from Sigma-Aldrich; palmitic acid was conjugated with BSA as described previously^{58 (link)}; ultrapure E. coli LPS serotype 0111:B4 (tlrl-3pelps) and Pam3CSK₄ (tlrl-pms) from InvivoGen; recombinant mouse IL-1Ra protein (HY-P72566) from MedChemExpress; recombinant mouse IL-6 protein (RMIL6I) from Invitrogen; recombinant mouse S100A9 protein (2065-s9), recombinant human S100A9 protein (9254-s9), recombinant human IL-6 protein (7270-IL/CF) from R&D systems; Pyruvate (L064-100) from Biowest; 2-deoxy-D-glucose (2-DG) (14325), ubiquitin isopeptidase inhibitor I G5 (21006), PR-619 (16276) and b-AP15 (11324) from Cayman Chemical Company. Acetonitrile and water 0.1% (v/v) formic acid were from J,T. Baker, and formic acid was obtained from Panreac. Ultrapure water filter through Milli-Q system (Millipore Corp).