Fig. 1.

Intestinal lamina propria CD11b⁺/CD11C⁺ mononuclear phagocytes from Nlrp3 ^R258W mice contain hyperactive inflammasome. a Colonic lamina propria mononuclear cells (LPMCs) were isolated from control mice (gray filled) or Nlrp3 ^eGFP reporter mice left untreated (blue line), or stimulated with heat-inactivated fecal bacteria (BAC) at multiplicity of infection (MOI) = 20:1 for 4 h (red line); different cell populations were identified by the indicated surface markers and were examined for NLRP3-eGFP expression. MFI represents geomean fluorescence intensity of eGFP in each sample. b, c Colonic LPMCs were stimulated with BAC (MOI = 20:1) for 12 h (b) or 5 h (c) with or without a ATP pulse for 30 min; control cells (MOCK) were not stimulated. IL-1β, IL-18, and TNF-α in the supernatants were determined through ELISA. The data are representative of at least three independent experiments and are shown as the means ± SEM. **P < 0.01, ***P < 0.001 (Two-tailed Student’s t test) (b); cleaved caspase-1 p10 subunit, IL-1β-p17 fragment in the supernatants (Sup), and their pro-forms together with NLRP3, ASC, and GAPDH in the cell lysates (Lys) were detected by western blot (c)