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. 2020 Apr 2;10:5811. doi: 10.1038/s41598-020-62602-x

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Cell death is linked to mitochondrial mass depletion in FECD CEC. (A) A marker of cell death by apoptosis (caspase-3/7 activity; green) was used in conjunction with the mitochondrial mass marker (mitotracker; red) in healthy and FECD explants. (B) Mitochondrial and caspase-3/7 signals were quantified in healthy (blue) and FECD (red) explants and plotted. Each dot represents a single cell. No caspase-3/7 signal was detected in healthy explants. Caspase-3/7 negative and positive cells composed 48% and 52% of the FECD cells, respectively. The median mitochondrial mass was higher in caspase-3/7 negative cells than in caspase-3 positive cells. FECD CECs with mitochondrial mass ranging between Caspase-3/7 positive and negative medians composed 46% of the FECD cellular population. These cells are referred to as containing a “Normal mitochondrial level”, which happened to be very similar to that of healthy cells. Twenty seven percent of the Caspase-3/7 negative FECD cells were above the median mitochondrial mass level and are therefore referred to as containing a “High mitochondrial level”. On the other hand, 27% of the caspase-3/7 positive FECD cells were below the median mitochondrial mass level and are therefore referred to as containing a “Low mitochondrial level”. Scale bar = 40 μm. Experiments were performed with 3 different explants from Healthy (900 cells analyzed, N = 3) and 3 FECD patients (846 cells analyzed, N = 3).