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. 2024 Jul 18;19(8):1172–1188. doi: 10.1016/j.stemcr.2024.06.006

Figure 5.

Figure 5

IL-22 activates JAK/STAT in human colonic epithelium organoids

(A) Bright-field images of control and IL-22-treated human colonic organoids for 3 days. Scale bars: 200 μm.

(B) Bar plot showing reduction of tdTomato+ cells (KRT20-tdTomato reporter line) following treatment with IL-22. Each dot represents an independent experiment (average of 3–6 replica wells per experiment). ∗p < 0.05 by paired Student’s t test.

(C) RT-qPCR analysis showing relative expression levels of indicated genes following IL-22 treatment. Plot shows fold change relative to untreated organoids ± SEM. Each dot represents an independent experiment. ∗p < 0.5 by two-way ANOVA.

(D) Representative images of human colonic organoids with IL-22 and/or JAK inhibitor tofacitinib for 3 days. Scale bars, 200 μm.

(E) RT-qPCR analysis of showing relative expression of the IA-associated genes following treatment with IL-22 and/or tofacitinib as in (D). Plot depicts fold change relative to untreated control organoids ±SEM; each dot represents an independent experiment. ∗∗∗∗ for p < 0.001 by paired two-way ANOVA followed by Dunnett’s multiple comparison test.

(F) Schematics of organoid formation assay setup. Bottom part shows representative images of organoids at 168 h formed following treatment with with IL-22 at the indicated concentrations during the initial 72 h.

(G) Time course RT-qPCR analysis confirming upregulation of REG1A upon IL-22 treatment. Pink shade indicates exposure window to IL-22. Each dot represents technical replicas of a single representative experiment.

(H) Quantification of organoid forming efficiency and organoid size following IL-22 treatment as indicated in (F). For efficiency, plot shows mean ± SD. Each dot represents a replica well of three independent experiments. For size, plot was generated using the Tukey method. The number of organoids analyzed within one representative experiment is shown below the bars. a.u, arbitrary unit. ∗∗, ∗∗∗, and ∗∗∗ for p < 0.01, 0.005, and 0.001, respectively, by two-way ANOVA.