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Salt washing of RNCs increases the accessibility of the attached nascent chains to MPP in vitro. Untreated and salt-washed RNCs were incubated in import buffer with 0.5 μM purified MPP in the presence or absence of 40 μg/ml purified NAC at 20°C for the indicated times. Cleavage of the Mdh1p precursor was analyzed by SDS-PAGE and fluorography. The percentage of cleaved chains was determined by densitometry of the fluorogram, with the amount of initially added Mdh1p set to 100%.
