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. 2024 Oct 8;43(22):5548–5585. doi: 10.1038/s44318-024-00242-x

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2024

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 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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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Figure EV1 — (A) Kernel density plots showing distribution of mitochondrial areas for indicated genotypes, measured from TEM images of heart mitochondria. Median values and, in parentheses, interquartile ranges are reported adjacent to curves. N = 2 animals per genotype except for Tfam mKO; Dele1 KO, where only 1 animal was available. >600 mitochondria were measured per animal. P value was <0.0001. Bar and error bars represent mean and SD, respectively. (B) Bar graph comparing the areas of segmented and non-segmented types of electrolucent mitochondria that were obtained from analysis of C10 G58R animals and littermates in (A). Statistics were performed using Mann–Whitney test, as the data had a non-parametric distribution. **** indicates p ≤ 0.0001. (C–E) Representative TEM images acquired at 2000× direct magnification show areas of myocardium of indicated genotype used for analysis of mitochondria. Scale bar = 5 µm. (F) Image of the subarea boxed yellow in (D), acquired at 5000× direct magnification and representative of the images used to quantify ultrastructural features of mitochondria detailed in Fig. 3. Scale bar = 2.5 µm. (G) Examples of inclusions observed in C10 G58R mutant mitochondria (black arrows). Scale bar = 500 nm. (H) Examples of two types of electrolucent mitochondria characterized by an enlarged matrix area absent of electron-dense substance and fewer cristae. (Top) A uniformly electrolucent mitochondrion. (Bottom) a segmented mitochondrion that has an electrolucent part (white arrow) separated from a portion of normal-looking matrix and cristae by a cut-through cristae. Open black arrowhead indicates the junction between electrolucent and normal portions of the segmented mitochondria. Scale bar = 500 nm. Please note: mitochondrion in (H, bottom) also appears in bottom right corner of image (J, top). (I) Mitochondria that are fully wrapped by electron-dense phagosome membranes (black arrows). Scale bar = 500 nm. (J) Mitochondria with ruptured OMMs. Open white arrowheads indicate sites where the intact IMM is visible, but OMM is absent. Scale bar = 500 nm. Please note: mitochondrion in image (H, bottom) also appears in the lower right corner of (J, top). Source data are available online for this figure.