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. 2017 Jun 13;7:3410. doi: 10.1038/s41598-017-03527-w

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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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Intracellular localization of TMHP and oxidized SOSG in the cells treated with CLD. (a) LCSM image of the cells treated with complete CLD in the serum-free culture medium for 1 h at 37 °C and fixed with 4% formaldehyde in PBS. Nuclei were stained with 0.3 µM DAPI. The left panel shows merged blue and red channels, the right panel shows differential interference contrast image. (b–g) Evidence for singlet oxygen generation due to PO-CL reaction in the cells. Confocal images of (b) intact cells, (c) the cells in the oxidative stress state, (d) similarly prepared cells treated with the complete and (e) reference CLD without TMHP. All samples were stained with 10 mM of SOSG (f) Distributions of the cells by the intensity of their fluorescence calculated from the analysis of at least 10 different images containing about 300 cells. 1 – intact cells, 2 – the cells under oxidative stress conditions, 3 – the cells prepared as in the sample (2) and treated with the complete CLD or 4 – with the reference dispersion without TMHP. (g) Measurement of SOSG endoperoxide fluorescence in the similarly prepared samples using plate fluorometer. Each sample contained about 0.2 millions of cells.