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. 2012 Jul 4;32(27):9143–9158. doi: 10.1523/JNEUROSCI.0416-12.2012

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Copyright © 2012 the authors 0270-6474/12/329143-16$15.00/0

This article is freely available online through the J Neurosci Open Choice option.

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Figure 7. — Validating the requirement of MtLS motifs for EB1-mediated MT plus end localization. A, Coimmunoprecipitation analyses in Cos-7 cells. Top, Extracts of Cos-7 cells expressing GFP-tagged Ctail, Ctail-3MtLS*, GRD-Ctail or GRD-Ctail-3MtLS*, before (lysate) or after (IP) enrichment through immunoprecipitation with GFP-Trap as indicated. Ctail-GFP and GRD-Ctail-GFP are expected to be 68 and 79 kDa in size, respectively, and a tendency of the protein to degrade could be suppressed to tolerable levels; middle, coimmunoprecipitation of endogenous EB1 is observed only in IP samples of Ctail (lane 3) and GRD-Ctail (lane 7), but not in the corresponding MtLS-mutant versions (lanes 4 and 8, respectively); bottom, anti-α-Vinculin reveals comparable amounts of loaded cell lysate extracts and also serves as an unspecific control for the coimmunoprecipitation. B, C, In primary Drosophila neurons, Ctail but not Ctail-3MtLS* tracks MT plus ends. Scale bar (in B) B, C, 5 μm.