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. 2021 Jan 26;4:119. doi: 10.1038/s42003-020-01642-5

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© The Author(s) 2021

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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Fig. 3 — A Schematic of EVs isolation and functional characterization. B Representative image of transmission electron microscopy (TEM) of cell line-derived EVs. C EV-surface markers in cell line-derived EVs by MACSPlex analysis. D miR-424 levels in EVs derived from RWPE-1 and LNCaP cells with (EVs-424) and without (EVs-CTRL) stable expression of miR-424. E Visualization of recipient cells incubated with fluorescently labeled EVs (PKH26, red) from RWPE-1 and LNCaP donor cells by confocal microscopy. Nuclei were stained with Hoechst (blue). F Level of miR-424 in RWPE-1 (upper) and LNCaP (lower) recipient cells incubated with EVs-CTRL or EVs-424. G Tumor-sphere formation by RWPE-1 (upper) and LNCaP (lower) cells supplemented with EVs-CTRL or EVs-424. H Confocal microscopy images of RWPE-1 recipient cells incubated for 24 h with EVs-CTRL or EVs-424 labeled with PKH67 (green) or PKH26 (red) fluorescent dyes. I Confocal microscopy images of tumor-spheres formed by RWPE-1 cells incubated with fluorescently labeled EVs as above. Right: quantification of fluorescently labeled tumor-spheres. J Growth of subcutaneous xenografts of RWPE-1 cells supplemented in vitro with EVs-CTRL and EVs-424 derived from stably expressing donor LNCaP cells. n = 4/group. K Tumor growth of LNCaP cells supplemented in vitro with EVs-CTRL and EVs-424 derived from stably expressing LNCaP donor cells. n = 4/group. Right panels, H&E and IHC staining for Ki67 in xenograft explants.*p ≤ 0.05, **p ≤ 0.01 by two-tailed Student’s t-test.