Figure 2

Effects of CAR on stemness gene expression and CSC sphere formation. (A) The U87MG cells were treated with DMSO (CTRL) or CAR (10 µM) in complete medium for 48 h. At the end of the incubation, a Real Time RT-PCR analysis of stem gene expression was performed. The data are presented as the mean values from three different experiments each performed in duplicate. (B–E) U87MG cells were incubated with DMSO or CAR (10 nM–20 µM) in a defined serum-free NSC medium for 9 days. (B) Representative image. The number of the newly formed spheres (C) and the mean diameter (D) were scored using the ImageJ program. The data represent the mean values from three pictures from two independent experiments. (E) Cells were treated as described above and a Real Time RT-PCR analysis of the stem cell markers or differentiated cell markers was performed. The data are presented as the fold of change vs. the expression at basal (day 0) level, before the treatment with the NSC medium, and are the mean values from three different experiments. The significance of the differences was determined by one-way ANOVA, followed by Bonferroni’s post hoc test: *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 vs. the CTRL; ^#P ≤ 0.05, ^##P ≤ 0.01, ^###P ≤ 0.001 vs. 0 day.