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. 2008 Dec 29;183(7):1213–1221. doi: 10.1083/jcb.200806048

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© 2008 Kutik et al.

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Figure 1. — Protein import and assembly defects in tam41Δ mitochondria. (A) Biogenesis stages I–V of mitochondrial carrier proteins (Ryan et al., 1999). (B) Isolated mitochondria from wild-type (WT) or tam41Δ yeast cells grown at 21°C were heat shocked in vitro and incubated with radiolabeled AAC or DIC precursor proteins at 25°C. The mitochondria were lysed with digitonin and analyzed by blue native electrophoresis and digital autoradiography. *, carrier form in tam41Δ mitochondria. (C) The assembly of AAC was analyzed as for B without in vitro heat shock. (D) After incubation with AAC, mitochondria were treated at pH 11.5. Total (T), pellet (P), and supernatant (S) were analyzed by SDS-PAGE followed by autoradiography and immunodecoration. (E) Isolated mitochondria (in vitro heat shock) were incubated with Fe/S protein. (F) Quantification of Fe/S protein maturation in the absence or presence of an in vitro heat shock in tam41Δ mitochondria. 20-min import into wild-type mitochondria was set to 100%. Error bars represent SEM (n = 3).