Fig. 5 |. iRHOM2 deficiency affects C. rodentium colitis and secretion of necessary cytokines in colon epithelial cells.

a, SourceTracker-based analysis of genera present in P2–4 stool. Likelihood of being sourced from gut, oral and skin microbiomes based on HMP data. b, Median c.f.u. per gram of stool for WT and Rhbdf2^−/− mice versus days of infection. c, Hematoxylin and eosin (H&E) stain and periodic acid–Schiff (PAS) stain of colon samples for WT and Rhbdf2^−/− mice at day 14 of C. rodentium infection. d, Flow cytometry dot plots quantitating surface mTNF expression on mice CD4 and CD8 T cells from lamina propria. e, Quantification of mTNF measured in d. f, Bead-based multiplex assay measurements of TNF in mouse serum during infection. g, Quantification of IL-6R measured in f. h, Percentage of CD4 and CD8 T cells in the lamina propia of WT and Rhbdf2^−/− mice expressing IFN-γ, IL-22, IL-17A and IL-17F following infectious colitis as quantified by flow cytometry (gating strategy is shown in Extended Data Fig. 6). i, Immunoblot of SW620 colon tumor epithelial cells with either iRHOM1 or iRHOM2 knockout. Asterisk, pro-ADAM17; black arrowhead, mature ADAM17. β-actin as endogenous control. j, Wound healing assay in iRHOM KO SW620 cells. Scratches were made and evaluated 72 h later. k, Quantification of wound healing measured in j (n = 4 per group). C.f.u. data were from three independent experiments, with five mice in each WT or KO group per experiment. For pathological findings, serum TNF and the cell phenotype of lamina propria were obtained from one (n = 10, respectively) out of three experiments. Two-way analysis of variance (b) or two-tailed, unpaired Student’s t-test (e,h,j; mean ± s.d.) or Mann–Whitney test (f; mean ± s.e.m.). Representative result of three (i) or five (c) independent experiments. nc, no change.