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. 2010 Dec 28;5(12):e15901. doi: 10.1371/journal.pone.0015901

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Wang et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Cumulus-oocyte complexes collected from control mice were stained with cytochrome c antibody and MitoTracker to determine the subcellular localization of cytochrome c (green) and mitochondria (red), and counterstained with DAPI to visualize nuclei (blue). Representative confocal sections of cumulus cells are shown. Cytochrome c shows a punctuate distribution pattern and co-localizes with the mitochondria (yellow) in control cumulus cells. (B) Cumulus-oocyte complexes collected from diabetic mice were stained with cytochrome c antibody (green), activated caspase-3 antibody (red) and DAPI (blue). All apoptotic cumulus cells of diabetic mice (arrows), as evidenced by positive staining of active caspase-3 antibody (red; arrows) and condensed chromatin (blue; arrows), always display diffuse staining of cytochrome c (green; arrows), indicating its translocation from mitochondria to cytoplasm. In contrast, those non-apoptotic cumulus cells of diabetic mice (green, lower panel; arrowheads) stained negatively with active caspase-3 antibody still show mitochondria-localized cytochrome c. Scale bars: 20 µm.