Rabbit anti caspase 1

Western blotting was routinely performed as previously reported [20 (link)]. Mouse anti-β-actin (1:10,000, Proteintech, Chicago, IL), rabbit anti-NLRP3 (1:500, Novus Biologicals, CO, USA), rabbit anti-caspase-1 (1:500, Proteintech), mouse anti-ASC (1:500, Santa Cruz, CA, USA), rabbit anti-GSDMD (1:1000, CST, Danvers, USA), mouse anti-pro-IL-1β (1:1000, Proteintech), rabbit anti-cleaved IL-1β (1:1000, Novus Biologicals), rabbit anti-pro-IL-18 (1:1000, Proteintech), rabbit anti-cleaved IL-18 (1:300, Bioss, Beijing, China), and rabbit anti-caspase-11 (1:1000, Novus Biologicals) were used. The densities of protein blots were quantified by using ImageJ software (NIH, Bethesda, MD) and normalized to the level of β-actin.

4-Hydroxytamoxifen was purchased from Aladdin (Shanghai, China). Jasplakinolide, taxol and caffeine were obtained from Abcam (Cambridge, UK). 2-Aminoethyl diphenylborinate, dantrolene, Z-VAD-FMK, SP600125, lipopolysaccharide (LPS) and nigericin were purchased from MedChemExpress (Monmouth Junction, NJ, USA). MCC950 were obtained from CSNpharm (Chicago, IL, USA). PEG8000 and PF431396 were obtained from Beyotime Biotechnology (Shanghai, China). Cantharidin was purchased from Sigma-Aldrich (Saint Louis, MO, USA). Rabbit anti-caspase-1, rabbit anti-ASC and rabbit anti-NLRP3 antibodies were obtained from Proteintech (Chicago, IL, USA). Mouse anti-α-tubulin antibody was from Boster (BM1452, Wuhan, China). Mouse anti-β-actin was purchased from Cell Signaling Technology (Danvers, MA, USA). The siRNA targeting ASC and CASPASE-1 was constructed by GenePhrama (Shanghai, China). The Flag-Gsdmd-NT, Flag-Gsdmd-CT plasmid were purchased from Addgene (Watertown, MA, USA).

All mice were sacrificed after behavior tests. Mice brains were fixed with 4% paraformaldehyde, and brain slices (4 μm thick) were prepared and used for staining. The slices were washed in PBS for 5 min three times and then blotted in 5% goat serum for 1 h at room temperature. The slices were incubated with the primary antibodies overnight at 4°C. The primary antibodies used were: rabbit anti-IBA-1 (1:200, Abcam, United Kingdom), rabbit anti-NLRP3 (1:200, Novus Biologicals, United States), rabbit anti-caspase-1, rabbit anti-GSDMD-N, and rabbit anti-NeuN (1:100, Proteintech, China). After being washed in PBS three times, these slices were incubated with secondary antibodies at 37°C for 1 h, and the secondary antibodies used were Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:400, Abcam, United Kingdom) and Alexa Fluor 594-conjugated goat anti-rabbit IgG (1:400, Abcam, United States). Then, these slices were washed three times again and stained with DAPI (Solarbio, China) for 5 min. The results were imaged using a fluorescence microscope (Nikon, Japan).