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. 2020 Sep 16;10:15160. doi: 10.1038/s41598-020-72122-3

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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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Immunomodulatory EVs induce elimination of tumor cells. In this assay B16- tumor cells were incubated with splenocytes and the indicated immunomodulatory EVs for 72 h, using 2 × 10¹⁰ EVs/well. Tumor cells were imaged by a high content imaging system (Operetta, Perkin Elmer) and the analyses were performed using the Columbus software 2.4.0 (Perkin Elmer, URL: https://www.perkinelmer.com/Product/image-data-storage-and-analysis-system-columbus). (A) B16F10, (B) B16F10 that were incubated with splenocytes, (C) B16F10 that were incubated with splenocytes and parental-EV, (D) B16F10 that were incubated with splenocytes and 41BBL-EVs, (E) B16F10 that were incubated with splenocytes and OX40L-EVs. One-way ANOVA statistical test followed by Dunnett’s test, *p < 0.05; mean and standard error. All comparisons were performed against the control of B16F10-cells.