Fig. 1

Generation and characterization of iPSC-derived neurons. a An induction method for the differentiation of neurons from iPSCs. b RT-qPCR analysis of stem cell marker (OCT4) and forebrain markers (FOXG1 and PAX6) compared between the original iPSCs (n = 3) and the induced neural progenitor cells (NPCs) at PID 15 (n = 3) (three independent differentiations). Bars, mean ± SEM. OCT4: ****p < 0.0001; FOXG1: ****p < 0.0001; PAX6: ***p = 0.0006 (Unpaired t-test). c Representative bright-field image of iPSC-derived neurons at PID 45. Scale bar; 100 μm. d Immunocytochemistry images of iPSC-derived neurons at PID 45 stained with neuronal markers (MAP2 and NeuN). Scale bar; 50 μm. e Relative proportion of MAP2 and NeuN expression compared to Hoechst of the differentiated neurons (n = 4) (four independent differentiations). Bars, mean ± SEM. f Immunocytochemistry images of synaptic markers (NR2B and Syn1). Scale bar; 10 μm. g Representative waveforms of neuronal activity (Ca²⁺ oscillations) of iPSC-derived neurons at PID 45 measured by Ca imaging using Fluo-8 and the treatment with 10 μM MK-801 (N-Methyl-D-aspartate (NMDA) receptor blocker) compared with control. h Quantification of Ca²⁺ oscillations (frequency (Spikes/Min) and amplitude (ΔF/F₀)) of iPSC-derived neurons at PID 45 measured by Ca imaging using Fluo-8 and the treatment with 10 μM MK-801 (n = 20) compared with the control (n = 20) (three independent experiments). Bars, mean ± SEM. ****p < 0.0001; **p = 0.0074 (Mann-Whitney test)