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. 2020 Jun 3;25(7):548–557. doi: 10.1007/s10495-020-01613-5

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© The Author(s) 2020

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 5 — Mitochondrial MMP, function and cellular ROS analysis. Jurkat cells were loaded with violet caspase, Zombie NIR, MitoTracker Deep Red and CellROX Green as described in the Materials and Methods section. Live (DN), early apoptotic (EAPO, Zombie NIR^+ve/caspase violet^+ve), late apoptotic (LAPO, Zombie NIR^+ve/caspase violet^+ve) and oncotic (Zombie NIR^+ve/caspase violet^−ve) cells were analysed for MMP by MitoTracker Deep Red (a), those MMP + ve cells analysed for CellROX Green, ROS (b), those MMP-ve cells analysed for CellROX Green, ROS (c). Untreated cells, 0.5 μM shikonin, blockade of shikonin with zVAD (20 μM) or blockade of shikonin with Nec-1 (60 μM) were analysed. Mean percentage of MFI was normalised to untreated cells ± SD, (n = 3) were analysed for significance by One Way ANOVA (P < 0.5) and post ad hoc tested for significance, NS (P > 0.5, not significant), significance was P < 0.05*, P < 0.01**, P < 0.001***, P < 0.0001**** compared to untreated cells and between treatments as indicated