Fig. 3

Relationship between LGR5 and reported CSCs markers and stem cell genes. a Analyses of CD133, CD44, CD90, CD24, and EpCAM expression in LGR5⁺ and LGR5⁻ U251 glioma cells and in LGR5⁺ and LGR5⁻ 8591 human primary glioma cells by FCM. b U251-GSCs and 8591-GSCs from U251 and 8591 human primary glioma cells through serum-free enrichment (top, scale bar = 100 μm) and immunofluorescence double staining for LGR5 and SOX2 (bottom, scale bar = 50 μm). c The expression levels of LGR5, CSCs markers (CD133, CD44, CD24 and EpCAM) and stem cell genes (SOX2, OCT4 and Nanog) in shLGR5 and shCtrl GSCs determined by Western blot (top). The protein intensity of these markers were determined by densitometric analysis and normalized to the relevant β-actin value. Error bars represent the mean ± SD of three independent experiments (bottom). *, P < 0.05; **, P < 0.01; ***, P < 0.001