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. 2000 Oct 2;151(1):29–40. doi: 10.1083/jcb.151.1.29

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© 2000 The Rockefeller University Press

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.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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The Arp2/3 complex binds to the NTA region of cortactin. (A) C3H 10T1/2 cell lysate (3 mg) was incubated with GST-, cortactin N-repeat 5–, cortactin NTA-, and cortactin-repeats–Sepharose beads for 2 h, washed, and associated proteins were analyzed by Western blotting with Arp3 antisera. C3H 10T1/2 lysate (18 μg) was loaded as a positive control (right). (B) In vivo association of the Arp2/3 complex with the cortactin NTA region. C3H 10T1/2 cells transfected with FLAG-full-length (FL), FLAG-N-term, FLAG-NTA, FLAG-repeats, and FLAG-C-term cortactin constructs were immunoprecipitated with anti-FLAG resin. Immune complexes were washed, subjected to 10% SDS-PAGE, and analyzed for Arp2/3 association by Western blotting with Arp3 antisera (top). The blot was subsequently stripped, and expression of FLAG-cortactin constructs was verified by blotting with M5 (bottom). C3H 10T1/2 lysate (20 μg) was loaded as a control (rightmost lane).