Fig. 5

Loss of NLRP12 in leukocytes triggers ISG induction within the intestinal epithelium. a Network analysis of differentially expressed genes (with log2 fc>1.5) in the caecum of Nlrp12^−/− mice. b Detection of STAT1 expression and activation by western blotting using total tissue samples from caecum of Nlrp12^−/− mice and wild-type controls (n = 3). β-Actin was used for loading control. c Top 5 up- and downregulated genes in the caecum of Nlrp12^−/− mice (p < 0.01) when compared to controls. d Validation of microarray-based gene regulation by RT-qPCR analysis (n = 5). e Relative gene expression of Ifi44, Ifit2, Oas2, and Apol9a/b was analyzed in the proximal colon of wild-type and mutant mice. Statistical significance was calculated by non-parametric Mann–Whitney test. P < 0.05 (*) was considered statistically significant. f Immunohistochemistry for IFIT2 was performed on 5 µm-thick tissue sections from caecum, proximal and distal colon of naïve wild-type and Nlrp12^−/− mice. Scale bars represent 50 µm (caecum) and 200 µm (proximal colon). g IFIT2 protein expression was analyzed by IHC on tissue sections prepared from proximal colon of wild-type chimeric mice (WT→WT), Nlrp12^−/− recipients that were reconstituted with hematopoietic cells from mutant mice (KO→KO), wild-type recipients that were reconstituted with hematopoietic cells from Nlrp12^−/− mice (KO→ WT), and Nlrp12^−/− mice that were reconstituted with hematopoietic cells from wild-type mice (WT→KO). Scale bars represent 200 µm. h Cytofluorometry analysis of living Ly6C^low MHCII^high (commonly referred as macrophages) that were isolated from the colon of chimeric mice (n = 6–8) following exclusion of NK cells, B cells, T cells, eosinophils, and neutrophils as previously described. Statistical significance was assessed by non-parametric Mann–Whitney test. P < 0.05 (*) and P < 0.01 (**) was considered statistically significant