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. 2017 Jul 24;7:6301. doi: 10.1038/s41598-017-06443-1

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© The Author(s) 2017

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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THP-1 cells respond to the CD63-negative EV fraction. (a,c) Pro-inflammatory THP-1 response depends on OMVs. 100 k pellets from L. pneumophila-infected or control THP-1 cells were generated. CD63-positive exosomes were removed by immunoprecipitation. Entire 100 k pellet or unbound EVs (CD63⁻ fraction) were used for stimulation of THP-1 cells for 24 and 48 h. Expression of CXCL8 (a), CXCL8 secretion (b) and expression of miR-146a (c) were analysed. Data are shown as mean + SEM of three independent experiments. ns: not significant.