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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 from NFAT3-expressing cells can further increase their anti-tumor effects and are effective in inhibiting tumor growth on a pre-established cancer. (A) 5.10⁵ MDA-MB-231 cells (D3H2LN-LUC) were injected into the left Fat Pad of each 6-weeks-old female Athymic Nude mouse. Tumor volume: Tumor growth is presented as the mean tumor volume (cm3) ±SEM, from mice injected weekly intra tumor with PBS or with EVs T-47D toCtl (5 × 10⁹ pp) or EVs T-47D ΔNFAT3, 7 days after cell xenotransplantation, 10 mice per group. Metastases: Beginning day 35, weekly, bioluminescent images (where the primary tumor was shield with a black tissue) were acquired on the IVIS system to quantify the mean photon flux produced by the metastatic cells. Metastases quantification is presented as the mean photon flux produced by the metastatic cells. Data from one representative experiment of two independent experiments is shown, all data are shown as mean ± SEM (n = 10 technical replicates;, *p < 0.05 **p < 0.005 and ***p < 0.001 compared to PBS-injected mice; (**), (***) compared to EVs-T-47D toCtl-injected mice). (B). 10⁵ MDA-MB-231 cells (D3H2LN-LUC) were injected into the left Fat Pad of each 6-weeks-old female Athymic Nude mouse. Tumor volume: Tumor growth is presented as the mean tumor volume (cm3) ±SEM, from mice injected weekly intra-tumor with PBS or with 5 × 10⁹ pp EVs T-47D ΔNFAT3, 38 days after cell xenotransplantation, 10 mice in each group. Metastases: Beginning at day 45, weekly, bioluminescent images (where the primary tumor was shield with a black tissue) were acquired on the IVIS system to quantify the mean photon flux produced by the metastatic cells. Metastases quantification is presented as the mean photon flux produced by the metastatic cells. Data from one representative experiment of three independent experiments is shown, all data are shown as mean ± SEM (n = 10 technical replicates; ***p < 0.001, compared to the PBS treated mice group).