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. 2018 Sep 18;7(2):255–274. doi: 10.1016/j.jcmgh.2018.09.006

Figure 2.

Figure 2

IL22-induced increases in enteroid size require claudin-2 up-regulation. (A) Cldn2 mRNA expression in jejunal enteroids cultured in ENR without or with IL22 (1 ng/mL, Peprotech), JAK inhibitor (JAK inhibitor I, 0.5 μmol/L), or STAT3 inhibitor (stattic, 5 μmol/L) for 3 days. n = 5 in this representative experiment. (B and C) Bright-field images, cross-sectional area (n = 12–20), and number (n = 6) of jejunal enteroids after culture for 3 days with IL22 (1 ng/mL, PeproTech), JAK inhibitor I, or stattic. Bar, 100 μm. *P < .05, **P < .01. (D) Bright-field images of jejunal enteroids from wild-type (WT), claudin-2 knockout (Cldn2 KO), and claudin-2 transgenic (Vil-Cldn2 Tg) mice cultured in ENR without or with IL22 (1 ng/mL, PeproTech) for 3 days. Bar, 200 μm. Images are representative of 3 independent experiments. (E and F) Cross-sectional area (n = 16–90) and number (n = 10) of jejunal enteroids derived from wild-type (WT), claudin-2 knockout (Cldn2 KO), and claudin-2 transgenic (Vil-Cldn2 Tg) mice after 3 days of culture with indicated IL22 concentrations. Data are representative of 3 independent experiments. **P < .01. (G) Morphologic analysis of EdU (red) incorporation into enteroids derived from wild-type (WT), claudin-2 knockout (Cldn2 KO), and claudin-2 transgenic (Vil-Cldn2 Tg) mice during 2-hour EdU pulse after 3 days of culture without or with IL22 (5 ng/mL). Bar, 100 μm. Representative data are shown.