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

Total RNA was isolated from cell cultures, and real-time PCR reactions were performed as described in Materials and Methods. Quantitative analysis of the relative expression of (A) SSEA-1, (B) Dcx, (C) β3-tubulin and (D) 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).