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. 2016 Oct 26;590(23):4147–4158. doi: 10.1002/1873-3468.12450

Figure 2.

Figure 2

TIM29 is a mitochondrial inner membrane protein, which forms a stable complex with TIM22. (A) Submitochondrial localization analysis using mitochondria or mitoplasts from HEK293T cells. Mito., mitochondria; MP, mitoplasts; Sonic., sonication; PK, proteinase K. Samples were analyzed by SDS/PAGE and western blotting. (B) Mitochondria were subjected to carbonate extraction at indicated pH or detergent lysis, followed by differential centrifugation; pellet, P; supernatant, S; total, T. Samples were analyzed by SDS/PAGE and western blotting. (C) Isolated mitochondria from HEK293T cells were solubilized in digitonin and both TIM22 and TIM29 were immunoprecipitated. Protein A‐Sepharose beads were used for the control column. Eluates were analyzed by SDS/PAGE, followed by western blotting using antiserum for the indicated proteins. Total 1%, Eluate 100%. (D) Isolated mitochondria from HEK293T cells were solubilized in digitonin and subjected to BN‐PAGE analysis (4–13% gradient), followed by western blotting and immunodetection.