Fig. 4

Establishment of neural differentiation platform of mouse embryonic stem cells. A Schematic of the ESC-to-NSC and NSC-to-neural differentiation transition processes. B Neurosphere formation during ESC-to-NSC transition and NSC-to-neural differentiation transition in 46C ES cells. Neurosphere size and number increased from day 1 to days 4 and 7. Scale bar, 100 µm. C qPCR for mRNA expression of ESC markers (Oct4 and Sox2), NSC markers (Sox1 and Nestin) and neuronal markers (Tuj1 and Map2). Data are presented as means ± SEM (n = 3). Significant difference and P values were analysed by one-way ANOVA with the Tukey’s multiple comparisons test. D Immunocytochemical analyses for the neural stem cell marker NESTIN and the neuronal marker MAP2 after neural differentiation for 14 and 21 days. Scale bar , 100 µm