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. 2023 Jan 12;5(1):129–146. doi: 10.1038/s42255-022-00715-5

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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, Immunofluorescence images and 3D reconstruction showing presence of Tom20⁺ material in cardiomyocyte-derived exophers from hearts of septic Card^RED mice. b, Presence of Tom20⁺ material in cardiomyocyte-derived exophers from hearts of Card^RED mice at SS and 3 d after CLP. Graph showing exopher numbers per field of view (FOV); n = 6 per group. c, TEM image of a mononuclear cell taking up mitochondria by extended pseudopods in septic hearts (CLP 7 d). d, Images showing cardiomyocyte-derived exophers (red) containing mitochondria (Tom20, white) present in TREM2⁺ macrophages (green) from hearts of Card^RED mice (n = 4). e, Cardiomyocyte-derived mitochondria (mt-Dendra2, green) present in TREM2⁺ macrophages (red) from hearts of MitoCard mice (n = 4). f, Cardiomyocyte-derived mitochondria (mt-Dendra2, green) localized in lysosomes (LAMP1, white) of TREM2⁺ macrophages (red) in hearts of MitoCard mice (n = 3). g, TREM2⁺ macrophages (green) took up cardiomyocyte-derived mitochondria (mt-Keima-458, cyan) and some mitochondria in an acidic environment (mt-Keima-561, red) from hearts of AAV9-Tnnt2-mt-Keima infected mice. h, CD163⁺ macrophages (green) engulfed cardiomyocyte-derived mitochondria in hearts of WT and Trem2^−/− mice infected with AAV9-Tnnt2-mt-Keima, respectively. Graph showing percentages of mt-Keima⁺ mitochondria in CD163⁺ macrophages (n = 5 per group). For each animal, we randomly selected five visualization areas and five Mac1 cells were analyzed. The ratio of the area of mt-Keima mitochondria in a single Mac1 cell to the area of the same Mac1 cell was calculated. The ratio in the KO group was normalized to the ratio of the control group. i, The incorporation of cardiomyocyte-derived tdTomato protein in CD45⁺CD11b⁺F4/80⁺CD163⁺RETNLA⁺ macrophages and CD45⁺CD11b⁺F4/80⁻LY6C⁺ monocytes from WT → Card^RED or KO → Card^RED chimeras 7 d after CLP. Graph showing quantification of MFI of tdTomato (n = 6–7 per group). j, Presence of Tom20⁺ material (cyan) in cardiomyocyte-derived exophers (red) from hearts of WT → Card^RED and KO → Card^RED chimeras 7 d after CLP. Graph showing the exopher numbers per FOV (n = 6 per group). Scale bars are indicated in the images. Bars show as mean ± s.e.m. (b,h,j) and median with interquartile range (i). Two-sided P values were determined by unpaired t-test (b,h,j) and Mann–Whitney U-test (i). Results represent four (b), three (d–f) and two (g–j) independent experiments (Extended Data Figs. 6 and 7).

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