Figure 4.
Effects of A1AR and A2BAR agonists on the expression of differentiation and stemness markers in CSCs. (a) CSCs derived from U87MG or U343MG cells were treated for 7 days with NSC medium containing DMSO (control), 100 nM CHA or 50 nM BAY606583. At the end of treatment periods, the total RNA was extracted, and relative quantification of the mRNAs for the stem cell marker CD133, the astrocyte marker GFAP, and the oligodendrocyte marker Olig2 were performed using RT-PCR, as described in the Materials and methods section. The data were expressed as the fold change versus the levels of the control and they are the mean values±S.E.M. of three different experiments. (b and c) CSCs from U343MG cells were treated as in a; at the end of treatment periods, cell lysates were subjected to western blot analysis using antibodies specific for the stem cell marker nestin or the glial-cell marker GFAP. (b) Representative western blots. (c) The bar graph shows the results of the quantitative analysis of the western blots, which was performed using the ImageJ program. The data were expressed as the percentage of optical density of the immunoreactive band relative to that of the control, set to 100% and are the mean values±S.E.M. of three different experiments. The significance of the differences was determined using a one-way ANOVA with the Bonferroni post-test: *P<0.05, **P<0.01, ***P<0.001 versus control