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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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L. pneumophila induced EVs elicit a pro-inflammatory response in non-infected THP-1 cells. (a,b) Response of THP-1 cells to vesicle-free supernatant or 100 k pellet from L. pneumophila-infected THP-1 cells. THP-1 cells were stimulated with vesicle-free supernatant or 100 k pellet from THP-1 cells infected with L. pneumophila (MOI 0.25 and 0.5, respectively) for 24 h. Recipient THP-1 cells were incubated for 24 or 48 h, respectively. (a) qPCR was performed for expression of CXCL8, miR-146a and pri-mir-146a. (b) Magnetic multiplex ELISA was performed. Results for TNF-α, IL-1β and MCP-1 are shown. Dashed lines indicate the input of the cytokine contained in the vesicle-free supernatant of infected THP-1 cells (MOI 0.5) before adding it to the recipient cells. Data are shown as mean + SEM of at least three independent experiments. *p < 0.05. *Compared to corresponding control, # compared to corresponding supernatant treated sample.