Fig. 2.

Atp6 processing is affected in INA22-mutant mitochondria. a Mitochondrial-encoded proteins were radiolabeled in vivo and analyzed by urea SDS-PAGE and digital autoradiography. b Wild type and ina22Δ mitochondria were solubilized in SDS-sample buffer, subjected to SDS-PAGE and Western blotting with indicated antibodies. c Wild type and ina22Δ mitochondria were subjected to in organello labeling for 20 min and protein co-immunoprecipitation with control or anti-Atp23 antibodies. Input (1%) and elution (100%) were analyzed by urea SDS-PAGE, digital autoradiography, and western blotting (WB) with anti-Atp23 antibodies. d Mitochondrial-encoded proteins were radiolabeled in vivo for 10 min and analyzed by urea SDS-PAGE and digital autoradiography or western blotting (WB) with indicated antibodies (left panel). Quantification was performed as in Figure 1b, n = 3, ±SEM. e Mitochondria were solubilized with 0.6% dodecyl maltoside (DDM) and analyzed by Blue-Native (BN)-PAGE followed by western blotting with anti-Atp5 antibodies (lanes 1–4), in-gel ATPase activity staining (complex V, lanes 5–8), or Coomassie staining (lanes 9–12)