Journal: Life Science Alliance
Article Title: Caspase-1 cleaves Bid to release mitochondrial SMAC and drive secondary necrosis in the absence of GSDMD
doi: 10.26508/lsa.202000735
Figure Lengend Snippet: Caspase-3 but not caspase-7 is required for lysis in Gsdmd −/− cel ls. (A) LDH release and caspase-3/-7 activity (DEVDase activity) from LPS-primed WT, Asc −/− , Casp1 −/− / Casp11 −/− , and Gsdmd −/− primary BMDMs after transfection of Salmonella typhimurium at MOI 10. (B) Immunoblots for caspase-3 and caspase-7 expression in lysates of Gsdmd −/− , Gsdmd −/− /Casp3 −/− , Gsdmd −/− /Casp7 −/− , and Gsdmd −/− /Casp3 −/− /Casp7 −/− iBMDMs. (C) PI influx from LPS-primed WT, Asc −/− , Casp1 −/− / Casp11 −/− , Gsdmd −/− , pool of Gsdmd −/− /Casp3 −/− clones (n = 3), pool of Gsdmd −/− /Casp7 −/− clones (n = 5), and Gsdmd −/− /Casp3 −/− /Casp7 −/− iBMDMs after transfection of poly(dA:dT). (D) Immunoblots showing caspase-3, caspase-7, and caspase-8 expression and LDH release after transfection poly(dA:dT) transfection from LPS-primed Gsdmd −/− iBMDMs transfected with control siRNA or siRNA targeting caspase-3, caspase-7, caspase-8 or both caspase-3/-7. Graphs show mean ± SD. ** P ≤ 0.01, *** P ≤ 0.001 (unpaired t test). Data and blot are representative of at least three independent experiments. Source data are available for this figure.
Article Snippet: VX-765 (MedChemExpress), Caspase-3/7 inhibitor I (CAS 220509-74-0; Santa Cruz Biotechnology), Q-VD-Oph (Selleck Chemicals), AZD5582 (Selleck Chemicals), 7-Cl-O-Nec1 (Abcam), GSK872 (Selleck Chemicals), K777 (AdipoGen), PD 150606 (Tocris), Calpeptin (Selleck Chemicals), ABT-737 (Selleck Chemicals), S63845 (Selleck Chemicals), and Nigericin (InvivoGen).
Techniques: Lysis, Activity Assay, Transfection, Western Blot, Expressing, Clone Assay, Control