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. 2017 Oct 31;7:14395. doi: 10.1038/s41598-017-14264-5

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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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Quantitative RT-PCR analysis of two miRNAs (miR-223-3p and let-7g-5p) identified by sequencing. The treatment of intact EVs with RNase before RNA extraction minimally altered Ct values of both miRNAs, strongly suggesting that these miRNAs were derived from the EVs-cargo and thereby protected from digestion by the membrane bilayer, whereas there was a large increase in Ct values when lysed EVs were similarly treated. Undetermined values (no detection by qRT-PCR) were assigned as Ct = 35. (A,B) Bar-graphs for miR-223-3p and let-7g-5p, showing Ct values for each treatment per sample; (C,D) Box-plots for miR-223-3p and let-7g-5p, showing Ct values per treatment (dots are results from each one of the five samples).