Fig. 1: IL-22 is over-expressed and ILC3s accumulate in the small bowel of Il10^−/− mice.

a IL-22 mRNA expression in distal, mid- and proximal colon and spleen of WT and Il10^−/− mice (n = 3–5). b IL-22 protein production in colon and small intestine explant supernatants from WT and Il10^−/− mice cultured for 24 h (n = 4–7). c Representative flow cytometry plots of IL-22 and IL-17A expression in CD4⁺ T cells in various tissues of the GI tracts of WT and Il10^−/− mice [MLN—mesenteric lymph nodes, cIEL—colonic intra-epithelial lymphocytes, cLPL—colonic lamina propria lymphocytes, sIEL—small intestine intra-epithelial lymphocytes, sLPL—small intestine lamina propria lymphocytes]. d Summary of IL-22-expressing CD4⁺ T cells in “superimposed” graph showing percentages of single positive (IL-22 SP) and double positive (IL-22/IL-17A DP) IL-22-producing cells from c. The total of all IL-22-producing CD4+ T cells is equal to IL-22 SP + IL-22/IL-17A DP, but not depicted on the graph. Data represent means from 3 to 5 individual experiments composed of cells obtained from 1 to 3 mice (males 20 weeks of age). Statistical differences in the percentages of IL-22⁺ T cells subsets between WT compared to Il10^−/− mice for each tissue are indicated as follows: a total IL-22⁺; b IL-22 SP and c IL-22/IL-17A DP. Statistical analyses (*p < 0.05, **p < 0.01 using the Mann–Whitney U test—error bars represent SD). e Representative flow cytometry plots of IL-22 and IL-17A expression gated on live singlet, CD3⁻ cells from the cLPL (top) and sLPL bottom of WT (left) and Il10^−/− (right) mice. f Summary of total ILC3 percentage in the indicated tissues based on the gating strategy in Supplementary Fig. 1. g Representative flow cytometry plots of IL-22 and IL-17A expression in ILC3s from the sLPL of WT (left) and Il10^−/− (right) mice based on the gating strategy in Supplementary Fig. 1. h Percentages of IL-22-expresssing (IL-22 SP) or IL-17A-expressing (IL-17A SP) ILC3s from the sLPL of WT (left) and Il10^−/− (right) mice as represented in g. I Serum IL-22 levels in WT and Il10^−/− mice (n = 10). Statistical analyses (*p < 0.05 **p < 0.01 using the Mann–Whitney U test—error bars represent SD).