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. 2018 Nov 19;8:17027. doi: 10.1038/s41598-018-35391-7

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© The Author(s) 2018

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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(A) Determination of mitochondrial superoxide using MitoSox showing increased superoxide in hypoxic tenocytes with normoxic control. Images in the left column show nuclear staining with DAPI; the images in the middle column show expression of superoxide while the images in the right column show overlay of MitoSox with DAPI. Images were acquired at 20x magnification using CCD camera attached to the Olympus microscope. (B) The image shows quantification of the mitochondrial superoxide. The intensity of protein expression as observed through immunofluorescence was acquired and the mean fluorescence intensity (MFI) was normalized to 100 cells. The graphs represent MFI mean values with standard error. The statistical significance of each hypoxic groups vs normoxic groups are represented in the figure (n = 4; NS – non-significant, *P < 0.05, **P < 0.01 and ***P < 0.001).