a, Schematic: generation of DG-like neurons from BD iPSCs.
b, Immunostainings of iPSCs for TRA-1–60 and Nanog, neural rosettes and neural progenitor cells for SOX2 and Nestin, and neurons for MAP2 and TUJ1. c, Immunostainings of neurons labelled with VGLUT1, MAP2, Prox1::eGFP and GABA. Scale bars, 50 μm for b and c. d, Quantification of VGLUT1-positive glutamatergic neurons (normal, n = 8; BD, n = 12 lines), Prox1::eGFP-positive DG-like neurons (normal, n = 8; BD, n = 12 lines) and GABAergic neurons (normal, n = 4; BD, n = 12 lines). e, Immunostaining of dendritic glutamatergic synapses and axonal GABAergic synapses. Scale bar, 5μm. f, Quantification of glutamatergic and GABAergic synapse densities (VGLUT1: normal, n = 30 neurons from 8 lines; BD, n = 78 from 12 lines. GABA: normal, n = 30 from 6 lines; BD, n = 88 from 12 lines).
g, Heat map of differential gene expression in normal and BD neurons.
h, Bar graph summarizing differential expression of mitochondrial genes in BD and normal neurons. i, Schematic rationale of JC-1. j, JC-1 flow cytometry graphs showing that, as a control, CCCP diminishes neuronal MMP and that BD neurons have elevated MMP. k, Quantification of elevated MMP in BD neurons compared with normal (normal, n = 8 lines from 4 subjects; BD, n = 12 lines from 6 subjects). l, m, Neurons expressing DsRed2-mito and Prox1::eGFP. Scale bars, 50 μm (l) and 20 μm (m).
n, DsRed2-mito puncta sizes reduced in BD neurons. Identical symbols indicate same subject (normal, n = 29 cells from 8 lines; BD, n = 39 from 12 lines). Student’s t-test, *P < 0.05; **P < 0.001. Bars, mean ± s.e.m.