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. 2020 Jun 2;10:8964. doi: 10.1038/s41598-020-65844-x

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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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EVs produced by poorly aggressive luminal breast cancer cells exert anti-invasive properties in vitro on different types of cancer cells. (A) Highly invasive triple negative breast cancer cells MDA-MB-231 were serum starved for 24 h and left untreated or were treated the following day with 3 × 10⁸ pp/mL EVs isolated from by WT T-47D; from WT MDA-MB-231 or from 2 different female primary human dermal fibroblasts (FHN21, FHN32) and subjected to in vitro invasion assay for 6 h. Data from one representative experiment of two independent experiments is shown, all data are shown as mean ± SEM (n = 3 technical replicates; **p < 0.005). (B) Poorly invasive luminal breast cancer cells T-47D were serum starved for 24 h and left untreated or were treated the following day with 3 × 10⁸ pp/mL EVs produced by WT MDA-MB-231 subjected to in vitro invasion assay for 24 h. Data from one representative experiment of two independent experiments is shown, all data are shown as mean ± SEM (n = 3 technical replicates; ***p < 0.001, compared to the untreated cells). (C) Highly invasive triple negative breast cancer cells MDA-MB-231 (left panel) and SUM-159-PT (right panel) were serum starved for 24 h and left untreated or treated the following day with 3 × 10⁸ pp/mL EVs produced by WT T-47D or by WT MCF7 and subjected to an in vitro invasion assay for 6 h. Data from one representative experiment of two independent experiments is shown, all data are shown as mean ± SEM (n = 3 technical replicates; **p < 0.005, compared to the untreated cells). (D) Highly invasive melanoma (WM.266.4), glioblastoma (U87MG) and pancreatic cancer cells (BXPC3) were serum starved for 24 h and left untreated or were treated the following day with 3 × 10⁸ pp/mL EVs produced by WT T-47D and subjected to in vitro invasion assay for 6 h. Data from one representative experiment of two independent experiments is shown, all data are shown as mean ± SEM (n = 3 technical replicates; **p < 0.005, compared to the untreated cells). For all data, the invasion index is calculated as a proportion of the number of invasive cells in treated wells compared to the number of invasive cells in the control well (−) arbitrarily set to 1.