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. 2008 Sep 8;182(5):937–950. doi: 10.1083/jcb.200801152

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© 2008 Claypool 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. — The largest AAC2-containing complex requires the presence of respiratory supercomplexes and is stabilized by CL. (A) Endogenous AAC2 coIPs with the respiratory supercomplex. An anti-complex IV IP was performed on digitonin extracts from the indicated mitochondria. 1% of the starting material and final flow-thru versus 100% of the preclear and IP beads post-washing were immunoblotted for the indicated mitochondrial proteins. n = 3. (B) Endogenous respiratory subunits coIP with AAC2. As in (A), except that 0.5% of the starting material and final flow-thru were analyzed. n = 3. (C) AAC2 and respiratory supercomplexes interact in organello. The anti-AAC2 IP was as in (B), except the preclear was omitted and the amount of protein analyzed is expressed as per mitochondrial source. n = 3. (D) wt or rho– mitochondrial extracts were resolved by 2D BN/SDS-PAGE and immunoblotted for the indicated mitochondrial proteins. n = 3. (E) After IP of wt digitonin extracts with normal rabbit serum or the anti-complex IV antiserum, the concentrated nonbinding flow-thrus were resolved by 2D BN/SDS-PAGE and immunoblotted for the indicated mitochondrial proteins. n = 3.