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. 2016 Aug 9;25(20):4405–4418. doi: 10.1093/hmg/ddw270

Figure 5.

Figure 5.

In vitro and in vivo rescue of neurite initiation via Dcx knockdown in 14-3-3ε overexpressing neurons. (A) Images of control pCAGIG (left panel), 14-3-3ε overexpression (panel second from left), control + Dcx (middle panel), 14-3-3ε + scramble miRNA (panel second from right) or 14-3-3ε + Dcx-miRNA (right panel) expressing Neuro-2A cells 36 h after transfection followed by 2 h of serum deprivation to induce neurite outgrowth. (Scale: 20 µm) (B) Analysis of the percent of cells with neurites, the mean number of neurites protruding from the soma and mean neurite length. Note that the overexpression of Dcx mimics the morphological deficits seen in 14-3-3ε overexpressing cells. Also note that the knockdown of Dcx in 14-3-3ε overexpressing cells rescues neurite initiation as seen by the percent of cells with neurites and the mean number of neurites protruding from the soma. Scale: 20 µm, n = 100 cells per condition per analysis from three independent experiments. (C) Images of P15 coronal brain sections following IUE at E16.5 of control mCherry + Scramble miRNA, mCherry-14-3-3ε + scramble miRNA or mCherry-14-3-3ε + Dcx miRNA expression plasmids. Bottom panels show magnified images of the cortical plate. (Scale: 100 µm) (D) Mean number of primary neurites protruding from the soma indicating that the knockdown of Dcx in 14-3-3ε overexpressing neurons rescues neurite initiation. (E) Analysis of mean neurite length. n = 25 neurons from three independent experiments per condition.