Fig. 5: Decreased neurite outgrowth with DISC1 mutation can be mimicked with UNC5D downregulation and rescued with UNC5D upregulation.
From: Convergence of independent DISC1 mutations on impaired neurite growth via decreased UNC5D expression
a Day 9 iNs were immunostained for UNC5D, MAP2, and F-actin as shown, revealing UNC5D expression in neurites, dendritic spines, and growth cones. Scale bar = 20 μm. b–c UNC5D shRNAs were transduced into day 4 iNs. WTex8 day 4 iNs were dissociated and plated for IncuCyte neurite length analysis with media alone (vehicle), viral transduction vehicle (empty), or else transduced with empty virus or 4 shRNAs targeting different sequences within UNC5D. Longitudinal neurite length analysis is shown. d, e MUTex8 day 4 iNs were dissociated and transfected with either dCas9-VP64 + RFP (dCas9) or dCas9-VP64 + UNC5D sgRNA #2 + RFP (dCas9 + UNC5D sgRNA). RFP-positive neurite length was analyzed longitudinally with IncuCyte. e Neurite length analysis at 192 h after plating (DIV 9) shows significantly increased neurite length with transient transactivation of endogenous UNC5D expression with gRNA #2. n = 25 for each group. Statistics: one-way ANOVA, *p < 0.05, **p < 0.01, ***p < 0.001, ***p < 0.0001
