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. 2019 Dec 11;9:18907. doi: 10.1038/s41598-019-55329-x

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© The Author(s) 2019

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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τ_bound exhibits an inverse correlation with oxidative stress in HEK293 cells. (A) Using monochlorobimane (MCB) imaging, the levels of reduced glutathione (normalised to inner sulcus, IS, as previously³¹) in each cell type at 2 W, 1 M and 1Y time-points showed a minor correlation (R² = 0.6) with the corresponding enzyme-bound NAD(P)H fluorescence lifetime data, suggesting that the antioxidative capacity of a system contributes, in part, to the value of τ_bound measured. (B,C) Culturing cells for 24 hours in rotenone (200 nM) or BSO (100 μM) each caused a significant increase in oxidative stress, based on the rate of DHE oxidation. Overexpression of NAD kinase (NADK+) caused this quantity to decrease (*P < 0.05). (D) Changes in oxidative stress were mirrored by opposite changes in τ_bound (*P < 0.001). NADK+ fluorescence lifetime data was taken from previous work¹⁹.