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. 2016 Mar 15;27(6):941–953. doi: 10.1091/mbc.E14-08-1332

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© 2016 Guillabert-Gourgues, Jaspard-Vinassa, Bats, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 1: — Kif26b is expressed in ECs and interacts with the Dvl3/Daam1 complex. (A) Quantitative real-time PCR of Kif26b transcripts in HUVECs, HMVECs, HeLa, immortalized HMVECs, and HEK293T (HEK) cells. (B) Endogenous Kif26b appears to colocalize with MTs in ECs. HUVECs immunostained with the antibodies against Kif26b (green) and α-tubulin (red). Bottom, high-magnification images of MTs. Scale bar, 15 μm (top), 5 μm (bottom). (C) Confocal analysis of HUVECs transfected with plasmids coding for Dvl3 with a Myc tag and immunostained with anti-Kif26b (green), anti-Daam1 (red), or anti-Myc (blue) antibodies. Bottom, high-magnification images. Scale bar, 10 μm (top), 3 μm (bottom). (D) Extracts from HeLa cells transfected with each indicated vector were immunoprecipitated with antibody against V5 (for V5-tagged N-MD mutant), Kif26b, or nonspecific immunoglobulin. Immunoprecipitates (IPs) and lysates were then immunoblotted with anti-Myc (for Myc-tagged Kif26b), anti-Kif26b, anti-V5, anti-Daam1, and anti-Dvl3 antibodies. (E) HeLa cells cotransfected with Kif26b-Myc and Dvl3 were treated with either si Control or si Daam1. Lysates were then immunoprecipitated with anti-Kif26b. IPs and lysates were immunoblotted with anti-Myc, anti-Daam1, and anti Dvl3 antibodies.