cDNAs for mouse Nlrp3, pro-Casp1, pro-IL1β, ASC and Gsdmd were amplified from reverse-transcribed cDNAs of J774 cells. Nlrp3, pro-casp1, pro-IL1β, ASC and Gsdmd cDNAs were inserted into a modified pBOBi plasmid vector with N-terminal or C-terminal 3× Flag or 3× HA tag. Truncation or point mutation of Gsdmd was generated by standard PCR cloning strategy. All plasmids were DNA-sequenced.
Antibody for IL-1β (5129-100) was purchased from Biovision. Antibody for Caspase-1 p20 (clone 4B4) was a kind gift from VM Dixit (Genetech, USA). Antibody for NLRP3 (AG-20B-0006-C100) was purchased from Adipogen. Antibody for GSDMD was purchased from Proteintech. Mouse anti-HA (F-7), anti-GAPDH (6C5) and ASC (sc-22514-R) antibodies were purchased from Santa Cruz Biotechnology, Inc. Mouse anti-Flag M2 and anti-HA beads, and anti-β-Tubulin (T6199) antibodies were purchased from Sigma-Aldrich. Ultrapure LPS from E. coli O111:B4, PI and Cholera Toxin B subunit were purchased from Sigma-Aldrich. Nigericin was purchased from InvivoGen. zVAD was obtained from Calbiochem. Anthrax Lethal Factor and Protective Antigen were purchased from List Biological Laboratories, Inc. CytoTox 96 Non-Radioactive Cytotoxicity Assay kit was purchased from Promega.
Antibody for IL-1β (5129-100) was purchased from Biovision. Antibody for Caspase-1 p20 (clone 4B4) was a kind gift from VM Dixit (Genetech, USA). Antibody for NLRP3 (AG-20B-0006-C100) was purchased from Adipogen. Antibody for GSDMD was purchased from Proteintech. Mouse anti-HA (F-7), anti-GAPDH (6C5) and ASC (sc-22514-R) antibodies were purchased from Santa Cruz Biotechnology, Inc. Mouse anti-Flag M2 and anti-HA beads, and anti-β-Tubulin (T6199) antibodies were purchased from Sigma-Aldrich. Ultrapure LPS from E. coli O111:B4, PI and Cholera Toxin B subunit were purchased from Sigma-Aldrich. Nigericin was purchased from InvivoGen. zVAD was obtained from Calbiochem. Anthrax Lethal Factor and Protective Antigen were purchased from List Biological Laboratories, Inc. CytoTox 96 Non-Radioactive Cytotoxicity Assay kit was purchased from Promega.
