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DIABLO is localized to the intermembrane space of mitochondria. Mitochondria (100 μg of protein) were isolated from yeast cells expressing preDIABLO and incubated in the presence (+) or absence (-) of 1.5 μg of trypsin with (+) and without (-) first rupturing the outer membrane by osmotic shock. After separation by SDS-PAGE, samples were analyzed by immunoblotting with antibodies recognizing DIABLO, the intermembrane space protein Cytb2 and the matrix-located mtHSP70.
