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. 2018 Jul 4;7(1):1–17. doi: 10.1016/j.jcmgh.2018.06.008

Figure 5.

Figure 5

IL22 limits ISC expansion. (A) Representative immunohistochemistry for the proliferation marker KI67 (red) and nuclei (blue) with quantification of the proportion of KI67+ nuclei. Technical n = 10+ organoids; biological N = 3 mice. Scale bar: 100 μm. (B–D) Gene expression analysis of organoids after 6 days with or without 500 pmol/L IL22 for (B) ISC-associated genes including Lgr5, Olfm4, Ascl2, and Sox9; (C) Wnt signaling pathway-associated genes including Wnt3, Ctnnb1, and Axin2; and (D) Notch signaling pathway-associated genes including Notch1, Notch2, Dll1, Dll4, Hes1, and Atoh1. Technical replicate n = 3, biological N = 3 mice. Significance was calculated using an unpaired t test relative to the untreated control. (E) Percentage organoid increase in response to 0, 60, or 500 pmol/L IL22 at each passage compared with the number of organoids at initial plating at p0. Technical replicate n = 3, biological N = 3 mice. Significance was calculated using 1-way analysis of variance with Bonferroni correction at each time point in comparison with control. +P < .05 for 500 pmol/L IL22 compared with control at passage 1. *P < .05, **P < .01, ***P < .001, and ****P < .0001.