FIG. 4.

MN treatment causes mitochondrial fragmentation in human corneal cells. HCEnC-21T cells were treated with 50 μM MN for 0, 1, 2, 4, and 6 h (left panel), fixed, and stained using anticytochrome c antibody and DAPI to reveal mitochondrial morphology and the nucleus, respectively. (A, left panel) Transition from the typical tubular mitochondrial network of normal cells (control and 1 h MN) into vesicular punctiform mitochondria (2, 4, and 6 h 50 μM MN). (A, right panel) Shows cytochrome c release after 50 μM MN treatment. (B) Mitochondrial morphology of two representative FECD donor specimens (61 male and 64 female) and representative normal donor specimen (59 male), labeled with cytochrome c (red) and DAPI (blue). Fragmented mitochondria (B, lower image) and cytochrome c release were around guttae (*). Shown are representative images of at least three independent experiments. DAPI, 6′-diamidino-2-phenylindole. To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars