Figure 4.

Thioridazine and fluphenazine reduce the percentage of cancer stem-like cell side population and affect the viability of U87MG sphere cells. (a and b) The cancer stem-like cells side population was significantly decreased by treatment with thioridazine (10 μM) from 1.26 to 0.03% in GBM8401 cells and from 1.43 to 0.1% in U87MG cells, as determined by side population assay. (c) Quantification of GSCs gene expression with GBM8401 and U87MG sphere cells by TaqMan real-time qRT-PCR, including CD133, CD44 and Nestin. Bar graph represents the mean of triplicates±S.D. **P<0.01, ***P<0.005. U87MG sphere cells were treated with 10 and 20 μM thioridazine (d) and fluphenazine (e) for 24 h, respectively. The viability of the U87MG sphere cells was determined by counting cells after trypan blue staining. (f) Both sphere cells and parental cells of GBM8401 and U87MG were treated with thioridazine at 5 and 10 μM for 24 h, respectively. LC3-I and LC3-II were detected by western blotting. GAPDH was used as an internal control