Figure 5.

Sphere differentiation assay of Fzd9+ and Lgr5+ cells. (A) The same numbers of Fzd9+ cells and Lgr5+ progenitors were cultured in vitro for 10 days and allowed to differentiate. (B) Immunofluorescence images of differentiated spheres and individual cells outside the colonies formed from Fzd9+ cells and Lgr5+ progenitors. The large square images are single XY slices, the vertical red line shows the position of the orthogonal slice that is shown to the right of each panel, and the blue line on the orthogonal slice shows the level of the XY slice to the left. Myo7a was used as the HC marker, and EdU was used to label proliferating cells. Myo7a+/EdU+ cells are indicated by white arrows. Scale bar, 20 μm. (C,D) Quantification of Myo7a+ and EdU+ cells in spheres (C) and Myo7a+ cells in inside and outside colonies (D) formed from Fzd9+ cells and Lgr5+ progenitors. n = 3 for both Lgr5+ and Fzd9+ cells. n.s., not significant.