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. 2018 Jul 9;8:10340. doi: 10.1038/s41598-018-28655-9

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© The Author(s) 2018

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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Characterization of EVs by flow cytometry. (a) ApogeeMix beads used to calibrate a BD LSRII to identify location of exosomes based on size. (b) A representative forward and side scatter of BAL EVs. (c) Frequency of CD63⁺ EVs. Mann Whitney T test, * < 0.05. (d–f) Frequency of EVs positive for HLA-DR, CD54, or CD36, after gating on CD63⁺ EVs. Mann Whitney T test, * < 0.05. (g,h) Mean fluorescence intensities of each marker analyzed on BAL EVs. Black bars represent healthy subjects (including SHS-exposed), and red bars represent asthmatics (including SHS-exposed). Mann Whitney T test, * < 0.05. (k–n) Mean fluorescence intensities of each marker analyzed on BAL EVs for each study group. Solid black bars represent healthy subjects, open black bars represent healthy subjects exposed to SHS, solid red bars represent asthmatics, and open red bars represent asthmatics exposed to SHS. Mann Whitney T test, * < 0.05; (Healthy Subjects, n = 9; Asthmatic Subjects, n = 9).