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. 2010 Oct 4;191(1):141–154. doi: 10.1083/jcb.201007026

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© 2010 Mick 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.rupress.org/terms). 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 3. — Coa3 associates with newly synthesized Cox1. (A) Wild-type (WT) and Coa3^HA mitochondria were solubilized and analyzed by BN-PAGE and Western blotting. (B) Wild-type and mutant mitochondria were analyzed as in A. (C) Radiolabeled Shy1, Cox5a, and Cox14 were imported into isolated mitochondria containing Coa3^HA. Mitochondria were solubilized and incubated with Flag (control) or HA antibodies before separation by BN-PAGE and digital autoradiography. (D) IgG chromatography of isolated mitochondria from wild type, Mss51^TAP, or Cox14^TAP. Samples were analyzed by SDS-PAGE and Western blotting. The asterisk indicates a cross-reactive signal detected by Mss51 antiserum. (E) In organello translation of isolated Coa3^HA mitochondria was performed, and the sample was split and subjected to coimmunoprecipitations with anti-Flag (control) or anti-HA antibodies. Samples were analyzed by SDS-PAGE and digital autoradiography. (D and E) The amounts of protein loaded in the total samples correspond to 2% of the eluate.

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