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. 2018 May 8;8:7227. doi: 10.1038/s41598-018-25515-4

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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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Exosomes released from cultured MDDCs carry Notch4 protein. MDDCs (from n = 3 healthy donors) cultured for 5 days with exosome-depleted FBS were treated with LPS (100 ng/ml) for additional 48 hrs to induce maturation (top left panel). Exosomes were isolated from culture supernatants of mature MDDC; EV concentration and size distribution was measured before and after (48 hrs) LPS treatment, by nanoparticle tracking analysis (bottom panel). Protein extracted from mature MDDC whole cell lysates and exosomes (30 μg/lane) was immunoblotted for exosome markers, Notch4, and GAPDH (top right). Lanes 1–3 = whole cell lysates, lane 4 = PBS blank, lane 5–7 = mature MDDC derived exosomes. Blots are cropped and full-length blots are included in Supplementary Figure S7.