Figure 5
Effect of NHI-1 on the sphere-derived cell morphology.
The GSCs were treated with complete NSC medium containing DMSO (control) or the indicated concentrations of NHI-1 for 7 days. As indicated, the drug-containing media were replaced with fresh drug-free NSC medium at the end of treatments and the cells were cultured for additional 7 days. (a) Representative cell micrographs after 7 days of treatment and after 7 days of drug wash-out were shown. (b) The area of the culture plates occupied by the spheres and (c) the length of cellular processes were scored after 7 days of treatment and after 7 days of drug wash-out. The counts represent the mean values ± SEM of three independent experiments. The significance of differences was determined by one-way ANOVA and Bonferroni’s post hoc test: **P ≤ 0.01, ***P ≤ 0.001 vs. the control; ###P ≤ 0.001 vs. the GSCs treated for 7 days. (d) The GSCs were treated with complete NSC medium containing DMSO (control) or the indicated concentrations of NHI-1 for 7 days. At the end of treatments, the total RNA was extracted and the relative mRNA levels of the stem cell marker CD133, the neuronal marker MAP2 and the astrocyte marker GFAP were quantified by real time RT-PCR. The data were expressed as the fold change compared to the levels of the control, which were set to 1 and are the mean values ± SEM of three different experiments. The significance of the differences was determined by one-way ANOVA and Bonferroni’s post hoc test: *P ≤ 0.05, ***P ≤ 0.001 vs. the control.
