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Fig. 3. Import of CIC into mitochondria that lack components of the TOM complex. (A) ³⁵S-labeled pCIC and mCIC were synthesized in reticulocyte lysate and incubated for 10 minutes at 25°C with isolated yeast mitochondria lacking Tom5. Mitochondria of the corresponding wild type were used in parallel assays. The import reaction was stopped by treatment with 250 µg/ml proteinase K at 0°C. The mitochondria were isolated again and the mitochondrial proteins were separated by SDS-PAGE. The relative amounts of imported CIC were determined using a phosphoimager. (B) Import of ³⁵S-labeled pCIC and mCIC into mitochondria that lack Tom22, following the same protocol as described in (A). (C) Import of ³⁵S-labeled Su9-DHFR into isolated mitochondria of the yeast strain tom22-2. (D) Import of pCIC into tom22-2 mitochondria.

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