Figure 7. Effects by loss of CDC42 are mimicked by a loss of Scribble in disrupting epithelial polarity and ISC-TA cell fate.

Scribble het (villin-CreERT2;Scribble^flox/flox) and Scribble KO (villin-CreERT2;Scribble^flox/+) mice were injected with TAM once per day for 3 days, and then sacrificed 1 day after the third TAM injection.

(A–C) Representative images of immunofluorescent staining of duodenal sections: anti-Na⁺K⁺-ATPase; enlargements of villi (A’, B’, and C’) and crypt (A”, B”, and C”).

(D and E) Small intestinal crypts were isolated, lysed, and immunoprecipitated with (D) anti-YAP and (E) anti-Scribble, then western blotted for (D) Scribble and (E) MST1/2 and Lats1/2. (representative of two independent repeats)

(F) Representative images of H&E staining of duodenal sections; one crypt is circled in each image.

(G–K) Representative images of immunofluorescence staining of duodenal sections: anti-E-cad, anti-Na⁺K⁺-ATPase, anti-YAP, anti-Ki67, anti-Pcna, αE-catenin, and anti-Olfm4.

(L and M) qRT-PCR analysis of (L) ISC marker, TA cell marker, and YAP pathway, and (M) Wnt pathway in isolated small intestinal crypts. Data are mean ± SD. *p < 0.05, **p < 0.01; ns, not significant.

For (A)–(C), data are representative of at least three independent experiments. Scale bars, 50 μm. n = 4 for each genotype. For (D) and (E), data are representative of two independent experiments. Scale bars, 50 μm. n = 3 for each genotype. For (F)–(M), data are representative of at least three independent experiments. Scale bars, 50 μm. n = 3 for each genotype.