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. 2020 Nov 3;10:18936. doi: 10.1038/s41598-020-75850-8

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© The Author(s) 2020

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/.

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Trafficking of necrotic bodies to endosomes-lysosomes in macrophages and glia. (A–L) Example images of macrophages (J774A.1 cells) (A–D), OECs (E–H) and SCs (I–L) (red) with NBs inside endosomes/lysosomes (green, arrowheads). Necrotic cells were tagged with pHrodo STP dye (green) that only fluoresces in acidic pH (i.e. in intracellular endosomes/lysosomes); NBs not in endosomes/lysosomes do not fluoresce (examples shown by arrows in F–H). (M–O) Graphical representation of NB appearance within endosomes-lysosomes (pHrodo-tagged) between macrophages (M), OECs (N) and SCs (O). The Y-axis shows the number of NBs in endo/lysosomes (green fluorescence) co-localizing with cells (red fluorescence) i.e. internalized NBs. The number of pHrodo-tagged NBs co-localizing with cells at the various time-points were compared to background levels (time zero); stars shows time-points at which there was a significant difference from background *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.0001 (two-way ANOVA with Sidak’s multiple comparison test). Data represents mean ± SEM. n = 3 biological repeats × 3 technical replicates × 4 FOV (n = 300–400 cells/FOV).