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. 2007 Jun 7;26(13):3227–3237. doi: 10.1038/sj.emboj.7601748

Figure 3.

Figure 3

mtDNA-encoded complex I (CI) subunits associate with CIA30 at early stages in their assembly. (A) BN-PAGE analysis of mitochondria containing radiolabeled mtDNA-encoded subunits (0 h chase) immunodepleted with the αCIA30 antibody (lanes 2, 4 and 6). Preimmune serum was also used for control depletion (lanes 1, 3 and 5). Protein complexes containing radiolabeled mtDNA-encoded subunits were detected by autoradiography (lanes 1 and 2). After this, the lanes were subjected to Western blot analysis using antibodies against CIA30 (lanes 3 and 4) followed by antibodies against CI and complex II (CII) (lanes 5 and 6). (B) After radiolabeling, cells were chased for various times as indicated. Mitochondria were isolated, solubilized in Triton X-100 and incubated with protein A-Sepharose coupled to CIA30 antibodies or preimmune sera. Bound proteins were analyzed by SDS–PAGE and phosphorimaging (lanes 2–7). A labeled subunit profile (control) is shown in lane 1 and represents 16.5% of material used in each co-immunoprecipitation. The mtDNA-encoded subunits are indicated.