Effects of P2X7R agonists and antagonists on the progress of neural differentiation, studied on the gene expression level.

<p>Total RNA was isolated from cell cultures, and real-time PCR reactions were performed as described in Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096281#s2" target="_blank">Methods</a>. Quantitative analysis of the relative expression of (<b>A</b>) SSEA-1, (<b>B</b>) Dcx, (<b>C</b>) β3-tubulin and (<b>D</b>) nestin in E14Tg2A cell line were performed by real time-PCR where GAPDH expression was used as internal control for normalization of expression levels. ESC were treated with P2X7R agonists or inhibitors (10 µM Bz-ATP or 10 µM KN-62) respectively. Data were analyzed by the Two-Way ANOVA test followed by the Bonferroni post-hoc test. The experiments were performed three times in triplicate with (***p<0.001 compared to control).</p>