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. 2017 Jul 18;7:5704. doi: 10.1038/s41598-017-05571-y

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© The Author(s) 2017

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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Ratio of geometric mean intensity between levels of the ROS generation in the vital cells with or without H₂O₂ stimulation. The rates of production in ROS from three mutant cell lines and three control cell lines were analyzed by FC500MCL flow cytometer system (Beckman) using (a) DCFDA (20 mM) and (b) MitoSox (5 mM), either without (Red) or with (Black) H₂O₂ stimulation. The relative ratio of intensity (stimulated versus unstimulated with H₂O₂) was calculated. (c) Total ROS generation in the vital cells and (d) mitochondrial ROS generation. The average of three independent determinations for each cell lines is shown. Graph details and symbols are explained in the legend to Fig. 2.