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. 2017 Sep 12;7:11271. doi: 10.1038/s41598-017-11249-2

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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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SEC-elution profile of EV samples. Conditioned media from melanoma cells (A) and T-lymphoblasts (B) cultures were processed using size exclusion chromatography (SEC). The obtained fractions were analysed by flow cytometry using anti-CD63 antibody after coupling EVs with aldehyde-sulfate beads. Protein concentration was also measured by BCA assay for each fraction and plotted in the right y-axis. Fold changes of mean fluorescence intensity (MFI) relative to the isotype control are plotted in the left y-axis. (C) Beads incubated with EVs (left) or EVs in solution (right) were incubated with primary and fluorochrome-labelled secondary antibodies. The flow cytometer gate used to analyse the beads is depicted in the SSC/FCS dot-plot. (D) Those beads showing CD63 positive staining were analysed by transmission electron microscopy. Bar: 100 nm.