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. 2021 May 5;11:9529. doi: 10.1038/s41598-021-88897-y

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© The Author(s) 2021

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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Microvesicles (MVs) released from pneumolysin (PLY)-treated lung alveolar epithelial cells are taken up by neutrophils. Human neutrophils (PMNs) were incubated with Ctr- or PLY-MitoTracker Deep Red- MVs [MVs were isolated from MitoTracker Deep Red-labeled A549 cells after treatment with PLY (100 ng/ml) or PBS- control (Ctr) at 37 °C or 4 °C for 1 h. PMNs were then analyzed by FACS for MitoTracker Deep Red signal. (A) Representative histogram of live PMNs (CD11b positive/7-AAD negative) treated with unlabeled MVs, Ctr- or PLY- MVs, at 37 °C. (B) Quantification of the percentage of MitoTracker Deep Red positive PMNs, n = 4, *p < 0.0001 (Mann–Whitney test, unpaired). (C) Representative histogram of live PMNs (CD11b positive/7-AAD negative) treated with PLY-MVs at 37 °C or 4 °C.