Gsdmd antibody

The mouse tumor tissue was fixed using 4% paraformaldehyde, then embedded in paraffin and sliced into 4-μm thick sections. The paraffin was removed with a citrate buffer solution (pH 6.0) and the sections were soaked in 3% H₂O₂ for 25 min. Following blocking with goat serum for 1 h, the sections were incubated overnight at 4 °C with Ki67 antibody (1:500, Abcam, Cambridge, UK), GSDMD antibody (1:500, Abcam), and GSDME antibody (1:500, Abcam). On the following day, the sections were stained with HRP-labeled secondary antibody for 30 min. Then the sections were restained with hematoxylin, dehydrated in gradient ethanol, cleared with xylene, and sealed with neutral gum. Immunohistochemical images were obtained under a light microscope (Olympus), and cell proliferation rates were calculated using ImageJ software.

Dextran sulfate sodium salt (DSS) was purchased from MP Biomedicals (CA, USA), adenosine triphosphate (ATP) was obtained from In vivoGen Biotechnology (CA, USA), Methyl sulfoxide (DMSO), lipopolysaccharide (LPS), and thioglycolate were purchased from Sigma-Aldrich (MO, USA). Carbobenzoxy-Leu-Leu-leucinal (MG132), chloroquine (CQ), and cycloheximide (CHX) were purchased from Selleck (TX, USA). Dithiothreitol (DTT), ethylene diamine tetraacetic acid (EDTA), and primers used in this study were synthesized by Sangon Biotech (Shanghai, China). Antibodies against NLRP3, Caspase-1, and ASC were purchased from Adipogen Life Sciences (CA, USA). Antibody specific to IL-1β was obtained from Cell Signaling Technology. GSDMD antibody was purchased from Abcam (Cambridge, UK). The Flag-tag antibody was obtained from Genscript (Nanjing, China). β-actin and HA-tag antibodies were purchased from Proteintech (Wuhan, China).

The reagents used are as follows: Nrf2 antibody, Proteintech, lot number: 16396-1-AP; NLRP3 antibody, Abcam, lot number: ab263899; Caspase-1 antibody, Servicebio, lot number: GB11383; GSDMD antibody, Abcam, lot number: ab219800; β-actin antibody, Proteintech, lot number: 20536-1-AP; and TUNEL kit: Proteintech, lot number: PF00006.