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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. 5 — A Assessment of EVs release from human tumor xenografts. B Growth of subcutaneous tumor xenografts of parental (CTRL) and miR-424-expressing LNCaP (424) cells in NSG mice. n = 4/group. C miR-424 level in explants of CTRL and 424 tumor xenografts. D miR-424 level in EVs isolated from xenografts (n = 2/group) of parental (EVs-CTRL) and miR-424-expressing LNCaP (EVs-424) cells. E Tumor-sphere forming assay with RWPE-1-recipient cells incubated with EVs-CTRL and EVs-424 xenograft-derived EVs. F Cell migration by Boyden chamber assay with RWPE-1 cells incubated with PKH26-labeled EVs-CTRL and EVs-424 from tumor xenografts. Left, representative phase-contrast and fluorescence microscopy images of RWPE-1 cells. G Schematic plan to evaluate EVs release from transgenic mouse models and functional characterization. H miR-322 levels in prostatic tissue from wild type (WT) and ERG/PTEN transgenic mice. n = 3/group. I miR-322 content in EVs isolated from prostatic tissue from WT and ERG/PTEN mice. n = 3/group. J Confocal images of RWPE-1 cells incubated for 48 h with EVs derived from WT (EVs-WT) and ERG/PTEN (EVs-ERG/PTEN) murine prostates. EVs were labeled with PKH26 (red) and supplemented to recipient cells. EVs (red). Nuclei are stained with Hoechst (blue). K miR-322 level in recipient RWPE-1 cells incubated with EVs-WT or EVs-ERG/PTEN. L Tumor-sphere assay with RWPE-1 cells supplemented with EVs-WT or EVs-ERG/PTEN. M Schematic of the experimental plan to block miR-424 in recipient cells. N Level of miR-322 evaluated in recipient cells supplemented with EVs from the indicated GEM source, following blockade of miR-424/322 using LNA-CTRL and LNA-424. O SFE in RWPE-1 recipient cells transfected with LNA-CTRL and LNA-424 and subsequent supplementation with EVs from the indicated GEM sources. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.005 by two-tailed Student’s t-test.