Inactivation of E-Cadherin and PTEN Alters Mammary Epithelial Morphology and Growth In Vitro
(A) Western blot analyses of E-cadherin, PTEN, and p-AKT(Ser473)/AKT protein expression of untransduced and AdCre-transduced Cdh1F/F, PtenF/F, and Cdh1F/F;PtenF/F mammary organoids.
(B) Representative bright-field images of untransduced and AdCre-transduced Cdh1F/F, PtenF/F, and Cdh1F/F;PtenF/F mammary organoids at day 21 of organoid culture. The dashed circle depicts the surface size of AdCre-transduced Cdh1F/F organoids. Scale bar, 50 μm.
(C) Average measurements of surface size (n = 10 per genotype) of untransduced and AdCre-transduced Cdh1F/F, PtenF/F, and Cdh1F/F;PtenF/F organoids at days 7, 14, and 21 of organoid culture.
(D) Expression of Ki67 and CC3 measured by whole-mount immunostaining of AdCre-transduced Cdh1F/F, PtenF/F, and Cdh1F/F;PtenF/F organoids. Scale bars, 50 μm.
(E) Quantification of the fraction of Ki67- and CC3-positive cells over the total cell number (determined by Hoechst) in AdCre-transduced Cdh1F/F, PtenF/F, and Cdh1F/F;PtenF/F (n = 10 per genotype).
(F) Representative bright-field images of a Cdh1F/F;PtenF/F organoid displaying invasive protrusions of single rows of cells (arrows) 1 day after medium change. Scale bars, 25 μm.