Figure 6. IL-22 administration overcomes CS-mediated inhibition of epithelial proliferation ex vivo and in vivo.

(A) Representative Western blot of SI organoids treated with or without MP (10 μM) for 24 hours, followed by treatment with rmIL-22 (5 ng/mL) for 2 hours. (B–D) Representative images and frequency and size of SI organoids cultured with or without MP and rmIL-22 (0.5 ng/mL) for 5 days (n = 3–4 wells per group). Scale bars: 200 μm. (E and F) Representative images and size of human SI organoids cultured with or without MP (10 μM) and rhIL-22 (10 ng/mL) for 6 days (n = 6 wells per group). Scale bars: 1,000 μm. (G and H) qPCR to determine Cdkn1a and Ccna2 expression in organoids cultured with or without MP (10 μM) and rmIL-22 (1 ng/mL) for 3 days (n = 3 wells per group). (I and J) B6 mice treated or not with MP (2 mg/kg i.p. daily) with or without F-652 (100 μg/kg s.c. every other day). Representative images and ileal crypt frequency and height on day 7 (n = 8–12 independent sections per group). Scale bars: 50 μm. (K–M) WT B6 mice were treated or not with MP (2 mg/kg i.p. daily) with or without F-652 (100 μg/kg s.c. every other day), starting 72 hours after TBI. Representative images, ileal crypt frequency and height, and Ki67⁺ cell frequency (n = 8–23 independent sections per group) on day 7. Scale bars: 50 μm. (N–Q) B6-into-BALB/c transplantation of BM with or without T cells. Recipients were treated or not with MP (2 mg/kg i.p. daily) with or without F-652 (100 μg/kg s.c. every other day), starting on day 7 after BMT. Ileal crypt frequency and height and Ki67⁺ and Olfm4⁺ cell frequency (n = 14–31 independent sections per group) on day 14 after BMT. Scale bars: 50 μm. Data are representative of at least 2 independent experiments or were combined from 2 experiments (K–Q). *P < 0.05, **P < 0.01, and ***P < 0.001, by 2-tailed t test or 1-way ANOVA.