Figure 1

PSA^-/lo PCa stem-like cells, as well as androgen-independent PCa cell lines exhibit elevated FOXC2 expression and key properties defining the EMT/CSC phenotype. (a) The left panel shows FACS plots representing sorting of GFP⁺ (PSA⁺) and GFP^-/lo (PSA^-/lo) fractions from LNCaP cells. The right panels show morphology and GFP fluorescence of sorted cells. (b) qRT–PCR analyses for FOXC2, and key prostate epithelial differentiation- (PD), neuroendocrine-differentiation- (NE), EMT- and stem-cell (SC)-related markers on sorted PSA⁺ and PSA^-/lo fractions from LNCaP cells analyzed immediately after sorting. Y-axis represents fold change in HPRT-normalized mRNA expression (n=3; error bars indicate s.e.m.) (c) Immunoblotting for FOXC2 and other indicated markers on sorted PSA⁺ and PSA^-/lo fractions. (d) IF for FOXC2 and indicated markers in sorted PSA⁺ and PSA^-/lo cells (scale bar, 100 μm; DAPI: pseudo-colored to red). (e) qRT–PCR analyses for FOXC2 and other indicated markers in PC3 and DU145 PCa cells compared with that in LNCaP cells. Y-axis represents fold change in HPRT-normalized mRNA expression compared with that of LNCaP cells (n=3; error bars indicate s.e.m.). (f) Immunoblotting for FOXC2 and other indicated markers in the above cells (***P<0.001). (g) Representative FACS plots for CD44 (APC) and CD24 (PE) surface marker expression analyzed in LNCaP and DU145 cells. (h) Quantification of FACS analysis shown in d (n=3; error bars indicate s.e.m.). (i) Quantitation of prostospheres (n=5; error bars indicate s.e.m.; ***P<0.001).