FIGURE 3.

(A) Representative images of double-staining for NeuN (Red) and GFAP (green) in cortical neurons in primary culture at 10 DIV. (B) Ratio of NeuN- and GFAP-positive cells to total cells (Hoechst-positive cells) at 10 DIV. Images of random four areas (each area of 1.3 mm²) of each well were taken, and 1,195–1,320 cells were counted in each well. The scale bar shows 100 μm. Results are the means ± SD (n = 4 independent experiments). (C–E) Changes in the protein levels of DNMT1 (C), DNMT3a (D), and DNMT3b (E) with time in culture. These DNMTs and β-actin bands were scanned, and the scanned bands were normalized by DIV2 cells on the same blot. β-Actin was used as a loading control. Results are the means ± SD (n = 5–13 independent experiments).