Fig. 6

Pathogenicity of dopaminergic neurons differentiated from PRKN-mutated TH-GFP iPSC lines. a Immunofluorescence staining of cleaved caspase-3 and GFP in TH-GFP iPSC-derived neurons under CCCP treatment for 24 h indicated that PRKN-mutated dopaminergic neurons were susceptible to cell death under oxidative stress. Arrowheads represent cleaved caspase-3/GFP double-positive PRKN-mutated cells. The arrow represents a cleaved caspase-3 single-positive PRKN-mutated cell. “PRKN” represents PRKN-mutated patient. Scale bar, 20 µm. b Quantitative analysis of the proportion of cleaved caspase-3-positive cells in GFP-positive dopaminergic neurons or GFP-negative non-dopaminergic neurons, determined from immunofluorescence images (n = 10 images per iPSC line) as in (a). “PRKN” represents PRKN-mutated patient. Values are shown as the mean ± SEM. Statistical significance was evaluated using the two-way ANOVA with Šidák’s multiple comparisons test. **P < 0.01, ****P < 0.0001. There was no significant difference in the proportion of cleaved caspase-3-positive cells between control and PRKN-mutated GFP-negative cells (P = 0.9901). There was also no significant difference in the proportion of cleaved caspase-3-positive cells between GFP-negative and -positive cells in the control lines (P = 0.3822)