Ac yvad aom

LPS from Escherichia coli 0111:B4, the antibody against β-actin (A5441), SP600125, PD98059, and cholesterol-water soluble were obtained from Sigma-Aldrich. Antibodies against phospho-p38 MAPK (Thr180/Tyr182; 4511), p38 MAPK (9212), phospho-p44/42 MAPK (Thr202/Tyr204; 9101), p44/42 MAPK (4695), phospho-SAPK/JNK (Thr183/Tyr185; 4668), SAPK/JNK (9252), and IL-1β (12242) were purchased from Cell Signaling Technology. Antibodies against ASC (22514), and active caspase-1 (sc-514 and 515) were obtained from Santa Cruz Biotechnology, Inc. Specific ON-Target plus siRNAs against murine mRNAs were obtained from Thermo Fisher Scientific. Silencer Select siRNAs specific to decrease the expression of human LPIN2 mRNA and negative controls were purchased from Ambion. Nigericine, the specific caspase-1 inhibitor Ac-YVAD-AOM, and SB203582 were purchased from EMD Millipore, and monosodium urate and alum were obtained from InvivoGen. The general caspase inhibitor Z-VAD-FMK was from APExBIO. The rest of the reagents were purchased from Sigma-Aldrich.

The human macrophage THP-1 cell line was from the American Type Culture Collection (Cat# TIB-202, RRID: CVCL_0006). HEK-Blue™ TLR4 (Cat# hkb-mtlr4; RRID: CVCL_IM89) and RAW-Blue™ cells (Cat# Raw-sp; RRID: CVCL_X594), Pam3csK4, and anti-TLR2 (Cat# mab2-mtlr2; RRID: AB_1277471) were from InvivoGen (San Diego, USA). E. coli lipopolysaccharide (LPS) 055:B5, phorbol 12-myristate 13-acetate (PMA), nigericin, MCC950, cytochalasin D, monodansylcadaverine, N-acetyl-L-cysteine, ATP and A438079 were from Sigma–Aldrich (San Luis, USA). Ac-YVAD-AOM and mitoTEMPO were from Merck-Millipore (Burlington, USA). Proteinase K was from Roche (Basel, Switzerland), DNase I from Qiagen (Hilden, Germany), Uricase from Worthington (Lakewood, USA), anti-HMGB1 (Cat# 651,401; RRID: AB_10945159) from Biolegend (San Diego, USA), and recombinant human IL-18 binding protein (IL-18BP) from GeneScript (Piscataway, USA).