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. 2023 Jun 20;14:3659. doi: 10.1038/s41467-023-39436-y

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© The Author(s) 2023

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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Fig. 5 — a mNeon and ΔVIT parasites were treated with FAC and CellROX Deep Red fluorescence quantified using flow cytometry. FAC-treated parental cells had no significant effect on CellROX fluorescence, however, in ΔVIT parasites, CellROX was significantly (p = 0.004, one way ANOVA with Sidak correction) increased. Each point represents geometric mean fluorescence of over 10,000 cells from n = 4, ns not significant. Bars are at mean, ±SD. b Treatment with NAC (5 mM) rescued ΔVIT parasite hypersensitivity to FAC. Points are the mean of n = 3, ±SEM. c Graph showing the mean EC₅₀ for FAC of mNeon and ΔVIT parasites, with or without NAC treatment. Each point represents an individual experiment, bars at the mean of n = 3, ±SD. p values from two sided extra sum of square F test. d ΔVIT parasites had significantly higher catalase activity compared to the parental line. Each point represents an independent experiment, n = 3, bar at mean ± SD. p value from one way ANOVA with Sidak correction. e mNeon and ΔVIT parasites were treated as in (a), and MitoSOX fluorescence quantified. Overlapping histogram from a single experiment showing change in fluorescence distribution. f Percentage of cells MitoSOX positive from four independent experiments, bars at the mean ± SD. p value from one way ANOVA with Tukey correction. g Immunofluorescence showing localisation and expression of parental and ΔVIT parasites endogenously expressing MIT-HA. Scale bar 5 um. h Quantification of MIT-HA signal as a ratio of TOM40. Points represent individual vacuoles from n = 3, bar at mean ± SD. p value from one way ANOVA with Tukey correction. i Western blot confirming the increased expression of MIT-HA in the ΔVIT parasites compared to the parental line, IMC1 used as loading control. Representative of 4 independent experiments. j Quantification of MIT-HA expression, normalised to loading control. Bar at mean ± SD, n = 3 individual biological replicates. p value from two tailed one sample t test.