Figure 7

Analysis of sub-lineages of c78^f/f mammary glands and efficiency of conditional gene recombination.

(a) Representative flow cytometric analyses of virgin mammary glands in 78^f/f (top panel) and c78^f/f mice (bottom panel) for surface markers (CD24α and CD49f) that distinguish myoepithelial (MYO, red circle), mammary repopulating unit (MRU, blue circle) and luminal epithelial cells (green circle) enriched fractions from stroma cells (pink circle). (b) Percentage of MYO, MRU and luminal epithelial cells (luminal) fractions in 78^f/f and c78^f/f mammary epithelial cells (n = 3 per genotype). (c) Representative pictures of mammary transplant outgrowths from epithelial cells isolated from 78^f/f and c78^f/f glands. There were outgrowths of both groups (n = 3 per genotype). Scale bar shows 1 cm and is applicable to all sections. (d) Flow cytometric analysis of re-isolated epithelial cells from c78^f/f mammary transplant. Red dots: MYO; Blue dots: MRU; Green dots: luminal colony forming cells (CFC). (e) Genotyping of Grp78 knockout and floxed alleles in the MYO, MRU and CFC enriched fractions shown in panel d. The relative ratios of signal intensities for the non-recombined (floxed) vs. recombined (KO) Grp78 allele were shown on right. Cropped gels were shown and the full-length gels were indicated in Supplementary Fig. 2c.