Fig. 1

Lgr5 expression profiled in the mammary gland throughout development. a Mammary glands were isolated from Lgr5KI embryos at embryonic days 15 and 17 (E15 and E17) and from Lgr5KI virgin adults. Whole mounts were immunostained for GFP (green) and DAPI (blue), and analyzed on a confocal microscope. b Single-cell suspensions were prepared from the mammary glands of Lgr5KI mice at stages E15, E17, and adult and stained for DAPI and EpCAM. Viable (DAPI negative), epithelial (EpCAM positive) cells were then measured for GFP expression through FACS analysis. The percentage of cells that were GFP positive (green) was measured by designating a threshold above which the epithelial cells of the wild-type littermates (gray) measured zero. c Viable single cells were sorted for GFP expression using the parameters mentioned in Fig. 1b and analyzed for expression of EpCAM and CD49f