Fig. 1

Differentiation of human iPSCs to GABAergic neurons. (a) Neural induction and differentiation protocol. Shh, sonic hedgehog; PM, purmorphamine. (b) Phase-contrast images of iPSCs, neural progenitor cells (NPCs) and GABAergic neurons (day 30) derived from control. Scale bar, 50 μm (c) Quantitative gene-expression analysis of each neurodevelopmental stage marker (Oct3/4, pluripotent stem cell marker; Nestin, neural stem cell marker; GAD1/67, GABAergic neuron marker) using iPSCs, NPCs and GABAergic neurons (day 30) derived from control and PARK2 (PA and PB). Control is presented as one column combining and averaging two cell lines (WD39 and 201B7). Data are presented as means ± SEM. Statistical analysis was performed using one-way ANOVA with Bonferroni’s post-hoc test (n = 3–4), ***p < 0.001 compared to control. (d) Schematic representation of the dorsal-ventral markers in the telencephalon. LGE, lateral ganglionic eminence; MGE; medial ganglionic eminence; POA, preoptic area. (e) Quantitative gene-expression analysis of control- and PARK2 (PA and PB)-derived NPCs for the expression of dorsal-ventral markers in the telencephalon. (f) Immunocytochemical analysis for the GABAergic neuron marker GABA and the neuronal marker βIII-tubulin of control and PARK2 (PA and PB) iPSC-derived GABAergic neurons. Scale bar, 50 μm. (g) Quantification of the percentages of βIII-tubulin- and GABA-positive cells. Data are presented as means ± SEM. Statistical analysis was performed using one-way ANOVA with Bonferroni’s post-hoc test (n = 3)