Subpopulations selected for high endogenous Rad6B expression (ZsGreen
) show elevated intracellular β-catenin. (A) ZsGreen fluorescence imaging of R6B-Zshigh MDA-MB-231 cells. a' is ZsGreen fluorescence merged with DAPI. Magnification ×20. (B) Immunofluorescence staining of R6B-Zshigh MDA-MB-231 cells with anti-Rad6 or anti-β-catenin, and detected with appropriate Texas Red conjugated secondary antibody. Endogenous ZsGreen fluorescence was merged with Texas Red and DAPI. Original magnification, ×40. (C) Immunofluorescence staining of control, R6B-Zshigh and R6B-Zslow MDA-MB-231 cells with anti-Rad6 or anti-β-catenin antibodies. Proteins were detected with FITC (Rad6) or Texas Red (β-catenin) conjugated secondary antibodies. Original magnification, ×20.