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. 2019 Aug 1;9:11187. doi: 10.1038/s41598-019-47593-8

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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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Differential regulation of mitochondrial membrane potential (ΔΨ_m) and mass at the interface vs. center in the µTSA. (A) ΔΨ_m levels assessed by TMRM fluorescence on Day 4 MCF-7-BMSC µTSA before and after addition of 20 μM FCCP uncoupler. Scale bar: 500 μm; (B) Representative radial distribution of TMRM fluorescence in a MCF-7-BMSC µTSA on day 4 before and after FCCP treatment. The red dot (r = 0.52) on the no-treatment (NTX) curve indicates that the TMRM fluorescence to the right of the dot is significantly higher than that of all FCCP-treated samples; from N = 6 independent experiments (p < 0.05, Welch’s t-test); (C) µTSA stained for mitochondrial mass with anti-TOM20. Right panels: confocal scans of the edge and the center. Green: TOM20; purple: vimentin; blue: DAPI. Scale bars: 500μm in widefield (left) and 25μm in confocal (right). (D) Fold difference in TMRM and TOM20 fluorescence between the edge and the center. N = 3 independent experiments. P-values: ordinary one-way ANOVA.