Fig. 3

miR-504 attenuates GSC stemness activity. a Immunofluorescence showing tumor spheres comprised of CD133-positive GSCs (red, CD133; blue, DAPI; scale bars = 100 μm). b GSCs differentiated into GFAP-positive cells after serum induction (red, GFAP; blue, DAPI; scale bars = 100 μm). c qRT-PCR detection of miR-504 expression in GSC-U87 and GSC-1295 cells transfected with miR-504 overexpression and inhibition vectors. d Representative images of tumor spheres formed after 14-days culture (scale bars = 500 μm). e Western blot analysis of the levels of the representative stem cell markers. f–h miR-504 reduced GSC-U87 cell tumorigenicity in vivo. i Immunohistochemistry for CD133 and KLF4 in xenograft sections (scale bar = 50 μm). Data are the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001