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. 2022 Nov 19;15(4):971–984. doi: 10.1016/j.jcmgh.2022.11.004

Figure 7.

Figure 7

IL13 drives the DCS cell population expansion in colonoids. (A) Expression levels of DCS cell markers Reg4, Spink4, and Kit, and the HDP Retnlb were quantified in murine colonoids treated with 10 ng/mL IL13 or 100 ng/mL IL33 for 24 hours (n = 6–8 cultures per condition). Dashed line represents expression in phosphate-buffered saline (PBS) control. (B) Colonoids treated with 10 ng/mL IL13 were immunostained for Reg4 (red) and E-cadherin (white) and quantified to assess DCS cell numbers. n = 10–13 cultures per experiment. Each data point represents the average of an individual culture in an independent experiment. (C) Colonoids were primed for 24 hours with PBS or IL13 (10 ng/mL) to induce DCS cell development, then starved in growth factor–deficient culture media for 48 hours. Cultures were analyzed for expression of stem cell markers Lrig1 and Lgr5. (D) PBS or IL13 primed (24 h) and starved (48 h) cultures were re-fed with growth factor–replete media for 24 hours and analyzed by quantitative polymerase chain reaction (qPCR) for Lrig1 and Lgr5 induction. (E) Colonoids treated with RELMβ (100 ng/mL, 24 h) were analyzed by qPCR. Data were analyzed by 2-way analysis of variance with the Tukey post hoc test or 2-tailed Student t test. ∗P < .05, ∗∗P < .01. CON, control.