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

Figure 6.

Figure 6.

14-3-3ε overexpression prevents the invasion of EB3 comets into initiating primitive neurites and the invasion defect is rescued by Dcx knockdown. (A) Time-lapse live imaging of EB3 dynamics in Neuro-2A cells during neurite initiation. Left panels demonstrate a representative time-lapse montage of a control cell expressing pCAG-CFP + EB3-mEmerald where microtubule comets can be seen invading primitive neurite. The middle panels depict a cell overexpressing 14-3-3ε + EB3-mEmerald where comets are not seen entering primitive neurite. The right panels show a cell overexpressing 14-3-3ε + EB3-mCherry (shown in green) + Dcx-miRNA where EB3 comet invasion into the initiating neurite is rescued. The top panel in each column shows the entire cell morphology with the white squares indicating the region shown in the panels below. Edge determined by phase contrast imaging and outlined with a white dotted line. Scale: 5 µm. See Supplementary Movies 5-7. (B) Mean number of comets to reach the edge of forming neurites every 30 s showing a significant decrease in 14-3-3ε overexpressing cells. (C) Mean comet distance from the forming primitive neurite edge indicating that microtubule comets are unable to reach the edge of forming neurites in 14-3-3ε overexpressing cells. (D) Mean number of comets in the entire soma indicating a decrease in microtubule activity in 14-3-3ε overexpressing cells.