Figure 2.

Neuronal Differentiation and Neuronal Maturation of hiPSC-NS/PCs Treated with or without GSI

(A and B) hiPSC-NS/PCs (with or without GSI treatment) dissociated into single cells, seeded on coverglasses, and immunostained for Ki-67 (cell proliferation marker) and Nestin (a neural stem cell marker). The nuclei were stained with Hoechst 33258. The micrographs show representative staining results for each group. Scale bars, 50 μm.

(C and D) Histograms showing the percentage of Ki-67⁺ and Nestin⁺ cells (n = 5 independent experiments).

(E and F) Fourteen days after neuronal induction, the hiPSC-NS/PCs were immunostained for βIII-tubulin (a neuron marker), GFAP (an astrocyte marker), and CNPase (an oligodendrocyte marker). The nuclei were stained with Hoechst 33258. The micrographs show representative staining results for each group. Scale bars, 50 μm.

(G–I) Histograms showing the percentage of βIII-tubulin⁺, GFAP⁺, and CNPase⁺ cells (n = 5 independent experiments).

(J) Daily analyses of neuronal maturation using MEA, which measures the active electrodes in each group (n = 5 independent experiments).

^∗p < 0.05, ^∗∗p < 0.01, and not significant (N.S.) according to one-way ANOVA with the Tukey-Kramer test. Data are presented as means ± SEM.