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. 2020 Jun 10;10:9414. doi: 10.1038/s41598-020-66120-8

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© The Author(s) 2020

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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Extracellular miR-1260b embedded on MFS-derived EVs. (A) Negative stain transmission electron microscopy (TEM) image of MFS cell lines (NMFH-1 and NMFH-2)-derived EVs. The image shows small vesicles approximately 100–150 nm in diameter. Scale bar =100 nm. (B) Representative dynamic light scattering analysis of EVs obtained from NMFH-1 and NMFH-2. The graph reports the diameter (nm) of the particles (x-axis) versus intensity (Optical Density y-axis). Numbers on graphs are the EV average sizes of NMFH-1 and NMFH-2. The isolated EVs ranged in size peak of 109 nm and 87 nm. (C) Western blot analysis on purified EVs derived from NMFH-1 and NMFH-2 and parental cellular lysates. CD63, a conventional EV marker. Cytochrome-C (Cyt-C), a mitochondrial marker. Calnexin, an endoplasmic reticulum (ER) marker. Vimentin, a mesenchymal marker. Tubulin as internal control for Western blot analysis. Full-length blots are presented in Supplementary Fig. 3. (D) EV-miR-1260b levels normalized to cel-miR-39 in MFS cells and hMSCs. Data are mean ± S.D. (n = 3 in each group) **p < 0.01, ***p < 0.001; Student’s t test. (E) EV-miR-1260b levels normalized to cel-miR-39 in MFS cells treated with the presence of absence of cambinol (48 hours). Data are mean ± S.D. (n = 3 in each group). *p < 0.05, **p < 0.01; Student’s t test. (F) EV-miR-1260b levels normalized to cel-miR-39 in MFS cells treated with the presence of absence of RNase. Data are mean ± S.D. (n = 3 in each group). N.S., not significant; Student’s t test.