Cell proliferation and cell division are required for Crb3-Venus-Gαi1wt–induced defective cystogenesis. (A) Schematics of experimental design. Stable Tet-Off MDCK cells expressing Crb3-Venus-Gαi1wt were cultured as indicated. dox, doxycycline. (B) Representative images of cysts grown for 6 d under different experimental conditions as depicted in A. Actin was stained with rhodamine phalloidin (red). DNA was stained with Hoechst 33342 (blue). Single confocal sections through the middle region of the cysts are shown. Merged images are shown on the right. Bar, 50 µm. (C) Quantitation of cysts with single lumen from MDCK cells expressing Crb3-Venus-Gαi1wt at d 6 under different experimental conditions as depicted in A. Values are mean ± SD from three independent experiments; n > 100 cysts/experiment. *, P < 0.001; **, P < 0.01.