Figure 6.

Epithelial cell expression of Mbd2 is vital to limit colitis. VillinΔMbd2 or littermate Cre⁻ (WT) mice were co-housed and subjected to 2% DSS in drinking water, tissues were harvested 8 days post consecutive treatment. The severity of colitis in the mice was assessed by (A) weight loss following DSS treatment and (B) symptom score (calculated from daily assessment of weight loss, rectal bleeding and stool consistency). (C) colon sections obtained from DSS treated mice were stained with H&E, and (D) colitis severity was assessed blinded according to a DSS histo-pathology score. Colon lamina propria cells were isolated and the relative proportion and number of monocytes, neutrophils, eosinophils, macrophages, DC subsets (E,F) and T cells (J) were assessed by flow cytometry. Lamina propria cells were incubated for 3 h with 1 μl/ml Golgistop and (G) the proportion of monocyte, neutrophil, eosinophil, cDC1s (CD11b⁻CD103⁺), and macrophages that express IL-1β (H) the proportion of myeloid IL-1β⁺ cells, (I) and the total number of IL-1β⁺ cells was assessed by intracellular staining and flow cytometry. (J) Number of CD4⁺ and CD8α/β⁺ TCRα/β T cells as a proportion of CD45⁺ cells. (K) ifng mRNA levels derived from 1 cm sections of distal colon in DSS treated VillinCreΔMbd2 or littermate Cre⁻ mice determined by qPCR relative to gapdh. All graphs show least mean square ±SEM, n = 15–25 per group, analyzed by linear regression of six independent experiments, except (J,K) which are representative data from three independent experiments. *P < 0.05, **P < 0.01, ****P < 0.0001.