Fig. 8.

DGCR14 is associated with the downregulation of PERK expression. (a) Relative expression levels of DGCR14 in DGCR14 knockdown cells. *** p < 0.001. (b) Immunoblotting for PERK and β-actin using the negative target and DGCR14 knockdown cells. ‘Negative target’ means the use of the negative control RNAi. (c) Quantification of the PERK/β-actin ratio. Three independent experiments were performed. The bars represent means ± SEs. * p < 0.05, ** p < 0.01. The value of N was set to 1. (d) Quantification of the mRNA expression level of DGCR14 in iPSCs and dopaminergic neurons (Day 24) derived from healthy controls and 22q11.2DS patients. The bars represent means ± SEs. 22DS1_2 and 22DS2_1 in dopaminergic neurons: n = 4; others: n = 3. *** p < 0.001. (e) Immunoblotting for puromycin and β-actin in DGCR14 knockdown cells. (f) Quantification of puromycilated protein signal intensity in e. The value of N was set to 1. The bars represent means ± SEs. Each n = 4. ** p < 0.01, *** p < 0.001 vs N. (g) Immunoblotting for puromycin and β-actin in dopaminergic neurons treated with cycloheximide (CHX). (h) Representative images of immunostaining for βIII-tubulin using CHX-treated dopaminergic neurons. The yellow bar in the image was 100 μm. (i) Measurement of neurite length. The numbers indicated in/over the bars represent the number of counted cells. The bars represent means ± SEs. *** p < 0.001. (j) Graphic abstract.