Modulation of neuroectodermal differentiation of mouse ESC by P2X7R activity.

(A) Flow cytometry analysis of SSEA-1 and β3-tubulin protein expression in neural-differentiated ES cells. Cells were cultured and induced to differentiation as described in Materials and Methods. (B) Immunofluorescence assay of SSEA-1 and β3-tubulin expression of cells following 8 days of differentiation. Circled areas: 1. cells expressing only SSEA-1, 2. cells co-expressing SSEA-1 and β3-tubulin. Scale bar, 50 µm. (C) Flow cytometry analysis of SSEA-1-positive cells co-expressing or not β3-tubulin on day 8 of differentiation cultured in the presence of 10 µM Bz-ATP or 10 µM KN-62, respectively. (D) Percentage of β3-tubulin-positive cells differentiated in the absence or presence of 1 µM A438079 or 10 µM KN-62, as determined by flow cytometry. Statistical relevance was analyzed by the One-Way ANOVA test followed by the Bonferroni post-hoc test. Six independent experiments were performed (* p<0.05 compared to control data).