Fig. 1: Characterization of isogenic RELN-del iPSCs and neurons.

a The target sites of the CRISPR-sgRNAs used in this study. CRISPR-sgRNA#1 and #2 target the sense strand, whereas sgRNA#3 and #4 target the antisense strand. b Schematic illustration of the differentiation of dopaminergic (DA) neurons. c Quantitative real-time PCR was performed to measure RELN expression in neurospheres (day 21) and dopaminergic neurons (day 28). CON1: neurosphere n = 4, DA neuron n = 5. CON2: neurosphere and DA neuron n = 4. IgCON1(+/−): neurosphere n = 4, DA neuron n = 3. IgCON1(−/−): neurosphere n = 3, DA neuron n = 4. IgCON2(+/−): neurosphere n = 3, DA neuron n = 6. IgCON2(−/−): neurosphere n = 4, DA neuron n = 5. Bars represent the mean ± SD. **p < 0.01, ***p < 0.001 vs respective parental neurospheres. ^†p < 0.05, ^††p < 0.01 vs respective parental DA neurons. d Analysis of dopaminergic neuron differentiation efficiency by quantifying the ratio of TH+ to TUJ1+ cells at day 23 (n = 180 cells). Bars represent mean ± SD. e Representative images of early dopaminergic neurons (day 23 = 48 h after the start of induction) immunostained for TH and reelin. The white bar in the image represents 200 μm. f Immunoblotting for phosphorylated tyrosine DAB1 (p-YDAB1), total DAB1, and β-actin using DA neurons at day 28. The values of p-YDAB1/total DAB1 are as follows: CON1 = 100, IgCON1(+/−) = 84, and IgCON1(−/−) = 75. g The venn diagram containing the altered gene expression in IgCON2(+/−) (blue) or IgCON2(−/−) (red). h GO analysis identified enriched categories for the altered genes in both IgCON2 neurospheres. A list of the top 10 categories showing the smallest p-values