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. 2020 Oct;159(4):1328–1341.e3. doi: 10.1053/j.gastro.2020.06.012

Figure 2.

Figure 2

Mtg8 and Mtg16 are regulated by Notch signaling. (A, B) qRT-PCR analysis of WT mouse intestinal organoids treated with Notch inhibitor DAPT for 2 and 3 days. Data represent mean ± SEM from biologically independent small intestinal organoid isolations (n = 3). The experiment was performed twice. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, 2-sided t-test. (C) qRT-PCR analysis of intestinal epithelium from Villin CreER and Villin CreER;Rbpjfl/fl mice collected at indicated days after tamoxifen induction. Data represent mean ± SEM from biologically independent animals (n = 4 per group). Three independent experiments were performed. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, 2-way analysis of variance (ANOVA). (D) RNAscope brown staining of Mtg8, Mtg16, and Mtgr1 in intestinal tissue obtained from Villin CreER and Villin CreER;Rbpjfl/fl mice collected at day 4 post-tamoxifen induction. Arrows indicate Mtg8+ cells. Scale bars, 50 μm. (E) RNAscope duplex staining of Mtg16 (blue) and Lgr5 (red) in intestinal tissues of the indicated genotypes at day 6 post-tamoxifen induction. (F) qRT-PCR analysis of intestinal epithelium from WT mice collected at the indicated days after DBZ or vehicle treatment. Data represent mean ± SEM from biologically independent animals (n = 3 per group). ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, 2-way ANOVA. (G) qRT-PCR analysis of DAPT-treated Villin CreER and Villin CreER;Atoh1fl/fl organoids induced with 4-OHT. Data represent mean ± SEM. The experiment was performed 4 times. ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, compared with untreated control group; #P < .05, ##P < .01, compared with DAPT-treated control group, 2-sided t-test.