Analysis of cancer stem cell spheroids from U87 and U373 cell lines for expression of the CSC marker CD133 by immunocytochemistry, semi-quantitative and quantitative RT-PCR, immunoblot, and flow cytometry.

<p>a) Photomicrographs of monolayer and spheroid colonies (left panel, scale bar = 50μm, n = 3) and CD133 marker staining (right panel, scale bar = 100μm) as shown by immunocytochemistry in monolayer and spheroid colonies of U87 and U373 cell lines. b) Quantitative RT-PCR of CD133 mRNA expression of U87 and U373 cell lines grown in CSC medium. Results are the mean ± SD of three independent experiments performed in triplicates (* p value <0.05, ** p value <0.01, n = 3). ML = monolayer, CSC = cancer stem cells. c) Flow cytometry analysis of CD133 in U87 and U373 cells grown in CSC medium.