dAIF regulates intramitochondrial localization of the Drosophila orthologue of CHCHD3/MIC19. (A) A total cell lysate, cytosolic, and mitochondrial-enriched fractions from flight muscles isolated from wild-type (w1118) adult flies were analyzed by SDS-PAGE and immunoblotting with anti-VDAC, anti-CytC, anti-Hsp60, and anti-Hsp90 antibodies to assess mitochondrial purity. The doublet for cytochrome C denotes both the oxidized and reduced forms of the coenzyme. (B) Mitochondrial preparations from flight muscles isolated from wild-type and Dmef2-Gal4/UAS-dMic19^-RNAi flies 4 d after eclosure were analyzed by SDS-PAGE and immunoblotting. RNAi-mediated disruption of dMic19 significantly reduces the amount of MIC19 that localizes to the mitochondrion. (C) SDS-PAGE and immunoblotting of mitochondrial preparations from flight muscles isolated from adult flies with the genotypes shown 2 d after eclosure. Intramitochondrial accumulation of MIC19 is reduced in all instances where AIF expression is knocked down. (D) SDS-PAGE and immunoblotting of mitochondrial preparations from wild-type and Mhc-Gal4/UAS-dErv1^-RNAi flies 2 d after eclosure. Intramitochondrial accumulation of MIC19 is reduced.