Fig. 6

The Nlrp3 ^R258W gut microbiota is shaped by IL-1β-induced antimicrobial peptides. a, b Colonic LPMCs and epithelial cells (ECs) were isolated from WT and Nlrp3 ^R258W mice and assayed for the expression of the indicated genes via Q-PCR without stimulation. The data are shown as the means ± SEM. c, d Colonic LPMCs and ECs were isolated from naive (c n = 3 for each genotype) or 4 days 4% DSS-treated (d n = 4 for each genotype) WT, Nlrp3 ^−/− and Casp1/11 ^−/− mice, the cells were treated with BAC (MOI = 20:1) ± ATP, and the culture media was assayed for IL-1β and IL-18 secretion via ELISA. The data are shown as the means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001 (one-way ANOVA with Tukey’s post-hoc test). e Changes in body weight in WT (n = 8), Il1r1 ^−/− (n = 9), Nlrp3 ^R258W x IL1r1 ^−/− (n = 6), and Nlrp3 ^R258W (n = 5) mice during DSS-induced colitis. The data are shown as the means ± SEM, *P < 0.05, **P < 0.01 (one-way ANOVA with Tukey’s post-hoc test). f The microbiota WT-index was also calculated for mice in e before DSS treatment. The medians with interquartile ranges are shown. *P < 0.05, **P < 0.01 (Kruskal–Wallis test). g Untreated WT (n = 4) and Nlrp3 ^R258W (n = 2) mice colon tissue RNA was assayed by RNAseq analysis, selected AMPs were shown to be upregulated in the Nlrp3 ^R258W colon at varied extent. *P < 0.05, **P < 0.01 (statistical comparison was conducted using DESeq2, see “Methods” section). h AMPs expression in untreated colon tissue from WT, Nlrp3 ^R258W, Nlrp3 ^R258Wx IL1r1 ^−/− and IL1r1 ^−/− mice were analyzed by Q-PCR. AU arbitrary unit. The data are shown as the means ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001 (one-way ANOVA with Tukey’s post-hoc test). i Colon tissues from Il1r1 ^−/− and WT mice were stimulated with 50 ng/ml mIL-1β for 4 h. Fold changes of AMPs expression (IL-1β stimulation vs. untreated) in colon tissues are shown. *P < 0.05 (two-tailed paired t test)