Fig. 4.

NLRP3 Inflammasome is required for IL-1β production by Cg-stimulated macrophages. a Peritoneal macrophages harvested from naive WT, Nlrp3^−/−, Nlrc4^−/−, Pycard^−/− mice were stimulated with Cg (300 μg/ml) or medium. At indicated time points, the supernatants were collected for quantification of IL-1β by Elisa. b, c Representative western blotting for immature (p35) or active form (p17) of IL-1β in the supernatant of Cg-stimulated macrophages (24 h). Macrophages were harvested from WT, Nlrp3^−/− or Pycard^−/− mice. d, e Peritoneal macrophages harvested from naive WT, Nlrp3^−/− or Pycard^−/− mice were stimulated with Cg (300 μg/ml) or medium. After 3 h, the Il1b mRNA expression was determined by qPCR. f Representative western blotting for ASC (p25) expression in Cg- or medium-stimulated macrophages harvested from WT mice. g Representative images (confocal) and quantification of ASC specks (green dots) formation in Cg (24 h)-stimulated peritoneal macrophages harvested from ASC-mCitrine reporter mice. Cell nucleus were stained by DAPI (blue). Data are represent the mean ± SD of four independent experiments compared Control vs WT (Cg) or WT vs Knockout groups to determine the level of statistical significance (*p < 0.05; **p < 0.01; and ***p < 0.001; ns, not significant)