Fig. 7.

Serial selection of GFP⁺/CK5⁺ stem-like cells from T47D^K5pGFP. a Diagram depicting a shift in the number and intensity of GFP⁺ cells upon triplicate P4 treatment and FACS isolation of the GFP⁺ fractions (3xP^high). b Immunofluorescence for CK5 (red) and DAPI (blue) in serial sorted cell line in absence of P4 (lower panel, “3xP^high”), compared with parental control + 100 nM P4 for 24 h (parental (“par”), upper panel). Scale bar, 50 μm. c Immunofluorescence for CK5 (red), PR (green) and DAPI (blue) in serial sorted cell line in absence of P4. Scale bar, 50 μm. d Parental and serial sorted cells as in a were placed in mammosphere/3D-Matrigel culture (300 per well), and the number of spheres ≥50 μm measured by automated counter after 7 days. Graph of mean number of spheres per field for each condition. Boxes represent range and lines represent means. *p < 0.05, unpaired t test