Figure 4

Suppression of WDFY1 expression restores impaired dendritic development in p39-deficient neurons. (a) Developmental expression of WDFY1 in rat cortical neurons at 4, 7, 10 and 14 days in vitro (DIV). (b–e) Elevated WDFY1 protein expression impaired dendritic development in hippocampal neurons. Rat hippocampal neurons at 7 DIV were transfected with vector control or Wdfy1 cDNA construct, and dendritic morphology was examined at 14 DIV. (b) Representative images showing the dendritic morphology of WDFY1-overexpressing hippocampal neurons. (c–e) Quantification of dendrite number (c) and total dendrite length (d), and Sholl analysis (e). Scale bar: 20 µm; n = 18–20 neurons from 3 independent experiments; ***p < 0.001, Mann–Whitney U-test. (f–i) Suppression of Wdfy1 restored the dendritic morphology in p39-deficient hippocampal neurons. Rat hippocampal neurons at 7 DIV were transfected with pSUPER vector control and shRNA targeting p39, Wdfy1, or both, and dendritic morphology was examined at 14 DIV. (f) Representative images showing dendritic morphology. (g–i) Quantification of the number of dendrites (g) and total dendrite length (h), and Sholl analysis (i); n = 14–16 neurons from 3 independent experiments; **p < 0.01, ***p < 0.001 versus Vector; ^#p < 0.05, ^##p < 0.01, ^###p < 0.001 versus sh_p39; ^†p < 0.05 versus sh_Wdfy1; Kruskal–Wallis one-way ANOVA followed by Dunn’s test.