Fig. 7

Treatment with carbonyl cyanide m-chlorophenylhydrazone (CCCP) and analysis of EV content. A A2780 cells were treated with CCCP (10 μm) and EVs were purified by immunoisolation. Mitochondrial protein expression was evaluated in EVs by Western blot analysis and B-H the relative abundance of RAB7, ATP5A, UQCRC2, SDHA, SDHB, NDUFS3, and NDUFB8 was determined through densitometric analysis normalizing against β-Actin. Parkin protein was used as control of CCCP treatment. I, J A2780 cells were treated with CCCP (10 μm) and EVs were purified by ultracentrifugation. RAB7 levels were determined, and their relative abundance was measured with densitometry normalizing against β-Actin. K A2780 CIS cells were transfected with empty vector (Mock) and with GFP-RAB7, and L OCR was determined with Seahorse Mito stress kit assay. M OCR and Extracellular Acidification Rate (ECAR) obtained with Seahorse instruments allowed to determine the energetic map. N-S Basal respiration, maximal respiration, ATP-production coupled respiration, proton leak, spare respiration capacity, and non-mitochondrial oxygen consumption were determined by Seahorse data elaboration. T-V A2780 CIS cells were transfected with GFP-RAB7, and DQBSA assay and LAMP-1 immunostaining were performed and analyzed by confocal microscopy. Data represent the mean ± SEM of at least three independent experiments. * p < 0.05; ** p < 0.01; ***p < 0.001