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. 2015 Feb 18;35(7):2942–2958. doi: 10.1523/JNEUROSCI.5029-13.2015

Figure 9.

Figure 9.

Depleting DBZ or expressing a phosphorylation mimetic form of Ndel1 results in a slower extension of EB3-EGFP, and coexpressing DISC1 rescues this phenotype. A, Using in utero electroporation gene transfer, the indicated vectors were transfected into the lateral ventricles of mouse embryos at E14.5. At E16.5, EB3-EGFP-expressing neurons were collected from the cortices and dissociated for primary culture. Forty-eight hours later, time-lapse images were obtained. Scale bar, 20 μm. EB3 binds to microtubule tips, and EB3 movements between the arrow and the arrowhead are shown in the kymographs. The results demonstrated that the EB3-EGFP velocity significantly decreased in DBZ-knockdown neurons. B, T219E:S251E Ndel1-expressing vectors or T219A:S251A Ndel1-expressing vectors were transfected together with the EB3-EGFP-expressing vector. The EB3 velocity in neurites significantly decreased in the T219E:S251E Ndel1-expressing neurons. C, D, The EB3 velocity in the neurites decreased in DBZ−/− neurons. E, F, Human DISC1 (hDISC1)-HA-expressing vectors were cotransfected into the lateral ventricle of mouse embryos at E14.5. Unlike DBZ−/− neurons without DISC1, no significant alteration in terms of EB3 velocity was observed among DISC1-overexpressing DBZ+/+ neurons, DISC1-overexpressing DBZ−/− neurons and neurons from WT. The values represent the mean ± SEM; *p < 0.05.