DAAM1 depletion causes extrusion of cells from spherical cysts. (A) Sphere cultures of shControl or shDAAM1 cells. Cells were cultured in Matrigel Matrix for 6 d. Control cells formed cysts with a radial cell arrangement, whereas DAAM1-depleted cells aggregated in disordered fashions with some cells being extruded from the sphere. Arrows indicate extruded cells. The results obtained using the shDAAM1-2 line are shown. Bar, 10 µm. (B) Quantification of cell extrusion from the sphere. Cells with a nucleus whose center stuck out of the sphere surface were regarded as extruded cells. Histograms represent the mean of three experiments. Error bars indicate SD. ***, P < 0.001.