Fig. 1

Differentiation and characterization of the cortical neurons derived from PD patients carrying SNCA A53T mutation. A Experimental timeline of differentiation and characterization of the iPSC-derived cortical neurons. Human neurogenin 2 (NGN2) is transiently overexpressed for 5 days for cortical neuronal induction. After induction of cortical neurons, analyses were performed from day 8 to day 15. B Representative images of MAP2 (green)-positive immunostaining in the differentiated neurons derived from the iPSCs on day 8 after neuronal induction. Scale bar = 100 μm. C Differentiation capacity of the differentiated neurons on day 8 after neuronal induction (n = 3 experimental replicates for each line; one-way ANOVA followed by Bonferroni’s multiple comparison test; N.S. not significant). D Representative images of α-synuclein (α-Syn) (red) in the differentiated neurons (green) on day 8 after neuronal induction. Scale bar = 50 μm. E, F Western blots of the differentiated neurons for α-Syn on day 8 after neuronal induction. β-actin (ACTB) was used as an internal control (n = 3 biological replicates; two-tailed Student’s t-test; N.S. not significant). G Quantification data of electrochemiluminescence assays of differentiated neurons for α-Syn on day 8 after neuronal induction (n = 3 biological replicates; two-tailed Student’s t-test; N.S. not significant). H Representative images of α-Syn-positive small aggregates (red) detected with anti-α-Syn oligomer specific antibodies in cortical neurons (green). Scale bar = 10 μm. I Quantification data of the number of α-Syn-positive small aggregates per neuron (n = 5 independent wells; two-tailed Student’s t-test; *p < 0.05). Bar graphs represent mean ± SD