Fig. 5

Role of eATP/P2X7R in the production of IL-1β by Cg-stimulated macrophages. a Peritoneal macrophages were maintained in regular medium or high-concentrated KCl (25 mM) medium and then stimulated by Cg (300 μg/ml). At indicated time points, the supernatants were collected for quantification of IL-1β by Elisa. b Peritoneal macrophages were pre-incubated with a selective inhibitor of P2X7R (iP2x7) (10 uM - 30 min) and then stimulated with Cg (300 μg/ml). After 6 h, the supernatants were collected for IL-1β quantification by Elisa. c Peritoneal macrophages harvested from naive WT or P2xr7^−/− mice were stimulated with Cg or medium. At indicated time points, the supernatants were collected for quantification of IL-1β by Elisa. d 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 or P2rx7^−/− mice. e Peritoneal macrophages harvested from naive WT or P2rx7^−/− mice were stimulated with Cg (300 μg/ml) or medium. After 3 h, the Il-1β mRNA expression was determined by qPCR. f Peritoneal macrophages harvested from naive WT mice were stimulated with Cg. At indicated times, the supernatants were collected for quantification of eATP concentration. g Peritoneal macrophages harvested from naive WT or P2xr7^−/− mice were stimulated with Cg or medium. At indicated times, the supernatants were collected for quantification of eATP concentration. Data are represent the mean ± SD of four independent experiments compared Control vs WT (Cg) or WT vs Knockout/Treatments groups to determine the level of statistical significance (*p < 0.05; **p < 0.01; and ***p < 0.001; ns, not significant)