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. 2014 Mar 3;204(5):839–856. doi: 10.1083/jcb.201306088

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© 2014 Shamir et al.

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Figure 1. — E-cad deletion induced loss of simple epithelial architecture. (A) E-cad deletion was induced in half of Cre-ER;E-cad^fl/fl organoids with tamoxifen. (B) Control, E-cad⁺ organoids (−Tam) formed cysts. (C) E-cad⁻ organoids (+Tam) failed to form cysts (28/42 movies across three biological replicates) or transiently established and then lost lumens (14/42 movies). (D) E-cad deletion blocked cyst formation. n, total number of organoids; r, number of biological replicates. Error bars indicate SD. ***, P = 0.0004, two-tailed Student’s t test with equal variance. (E) Control organoids formed cysts with enrichment of E-cad and ZO-1 along apicolateral membranes (E’). (F) E-cad⁻ organoids were multilayered, lacked E-cad immunoreactivity, and displayed abnormal ZO-1 localization. Arrow indicates rare E-cad⁺ cells. (G) By Western blot, E-cad deletion (+Tam) resulted in complete loss of E-cad protein and significant reductions in αE-catenin and β-catenin (see also Fig. S1, A and B). Whole cell lysate samples were loaded for equal protein based on BCA analysis. (H) The Cre biosensor mT/mG was used to observe E-cad⁻ cell behaviors by confocal microscopy (Video 1). Cre⁺, E-cad⁻ cells (green) changed shape, from columnar to round, before shifting apically (H’ and H’’, arrowheads). Gamma adjustments were performed in E and F to improve image clarity. Bars: (B and C) 20 µm; (E, F, and H) 10 µm.