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. 2008 May 5;181(3):537–549. doi: 10.1083/jcb.200707022

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© 2008 Kakinuma et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 4. — Kank inhibits the formation of actin stress fibers through 14-3-3. (A) Kank inhibits the formation of actin stress fibers. Kank or its mutants (Kank^Δcoil-FLAG, Kank with deletion of amino acids 90–360, and Kank^S167A-FLAG) were expressed with or without 14-3-3θ in NIH3T3 cells. The cells were incubated with an anti-Kank pAb or anti-Kank mAb together with an anti-GFP antibody. Kank protein was detected using Alexa Fluor 488–conjugated goat anti–rabbit IgG (green, top middle) or Cy5-conjugated goat anti–mouse antibody (purple, bottom middle). GFP protein was detected using Alexa Fluor 488–conjugated goat anti–rabbit IgG (green, left). F-actin was stained with TRITC-conjugated phalloidin. Arrowheads indicate the cells expressing the transfected vectors. Bar, 10 μm. (B) The number of the cells with actin stress fibers was quantified. Kank and its mutants were transiently expressed and the cells were stained as described in A. 100 cells expressing the transfected genes were examined and the results shown are the means ± SD of three independent experiments. *, P < 0.01 compared with the control; **, P < 0.05 compared with the control.