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. 2023 Jul 10;21:171. doi: 10.1186/s12964-023-01186-1

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© The Author(s) 2023

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Stimulation with EVs derived from ROR1/2-overexpressing cells increases tumor invasion dependent on RhoA. A + B Boyden chamber invasion assays of MCF-7 cells after stimulation with (A) EVs isolated from MCF-7 empty vector (pCMV/pcDNA) or ROR1/2-overexpressing (pROR1/pROR2) cells or (B) EVs from MDA-MB-231 CRISPR control (Cr-CTL) or ROR1 KO (Cr-ROR1) cells (mean ± SD, n = 3). C Boyden chamber invasion assays of MCF-7 cells transfected with siRNA against RhoA (10 nM) 24 h prior to stimulation with EVs isolated from MCF-7 empty vector (pCMV/pcDNA) or ROR1/2-overexpressing (pROR1/pROR2) cells (mean ± SD, n = 2). D Immunofluorescence: MCF-7 cells were pre-treated with/without dynasore (12.5 µM) for 2 h and uptake of PKH26-labeled EVs or dye-only (PKH-CTL) into the cells was visualized by confocal microscopy (n = 3). Signals were quantified with ImageJ. Boxplots depict the median (line), the 25–75 percentiles (box) and the 10–90 percentiles (whiskers) of n = 15 quantified fields. Scale bar: 10 µm. E Boyden chamber invasion assay of MCF-7 stimulated with pROR1/pROR2 EVs after pre-treatment of the cells with/without dynasore (12.5 µM) for 2 h (mean ± SD, n = 3)