Caspase 11 sc 374615

3-MA was purchased from Selleckchem (Houston, TX, USA). Antibodies to Beclin-1 (#3738), Atg7 (#2631), and p53 (#2527) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies to GAPDH (sc-32233), Collagen I (sc-28654), and Caspase-11 (sc-374615) were purchased from Santa Cruz Biotechnology (San Diego, CA, USA). Antibodies to NLRP3 (ab214185), and Caspase-1 (ab179515) were purchased from Abcam (Cambridge, MA, USA). Antibodies to Collagen III (GB11023), and IL-1β (GB11113) were purchased from Servicebio (Wuhan, China). Antibodies to IL-18 (A16737) and GSDMD (A20197) were purchased from ABclonal Biotech (Shanghai, China). Antibody to cathepsin B (12216-1-AP) was purchased from Proteintech Group (Chicago, IL, USA). Anti-mouse secondary antibody (A0216), and anti-rabbit secondary antibody (A0208) were purchased from Beyotime Institute of Biotechnology (Shanghai, China). Beclin-1 siRNA and GP-transfect-Mate (G04009) were purchased from GenePharma (Shanghai, China). Antibody to α-SMA (A2547), DMSO and all other chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Total protein from BMDM, neutrophils, lymphocytes and mentioned tissues was extracted in ice cold RIPA buffer (C-9806S, Cell Signaling Tech. Beverly, Massachusetts) containing protease inhibitors (P8340, Sigma Aldrich St. Louis, Missouri). Proteins were detected by immunoblotting using standard techniques. Antibodies that were used: caspase-11 (sc-374615), GAPDH (sc-47724), β-ACTIN (sc-47778), Pol II (sc-9001), GCN5 (sc-20698) from Santa Cruz; caspase-1(AG-20B-0042) from adipogen; anti-COMMD10 antibody (GTX121488) from Genetex, pP65 (3033) and P65 (6956) from Cell Signaling. Blots were incubated with HRP-conjugated secondary antibodies, and subjected to chemiluminescent detection using the MicroChemi imaging system (DNR Bio-Imaging Systems, Israel). P65 subcellular fractionation was performed using NE-PER nuclear/cytoplasmic extraction kit (78835, Thermo scientific, Paisley, UK) per manufacturer's instructions. Equivalent protein amounts were loaded for both nuclear and cytoplasmic fractions.