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. 2019 Jan 17;21:7. doi: 10.1186/s13058-018-1088-6

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© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 3 — Adipocytes increase the nuclear efflux of doxorubicin (DOX) and its expulsion from tumor cells via extracellular vesicles (EVs). a Left panel: intracellular localization of DOX visualized by confocal microscopy in non-cocultivated (NC) and cocultivated (C) cells at indicated times after DOX exposure in MDA-MB436 and E0771 cells. Nuclei were labeled with 4′,6-diamidino-2-phenylindole (DAPI) (scale bars, 10 μm). Right panel: corresponding quantification of fluorescence intensity (DOX) in the nuclei. b Analysis of the cytoplasmic localization of DOX in live MDA-MB436 cells using time-lapse video microscopy in control cells (NC) and cells cultivated with adipocyte-conditioned medium (AdCM) for both the pre- and post-incubation steps at indicated times. c Left panel: one representative analysis using nanoparticle tracking analysis (NTA) technology of the number of EVs secreted by MDAMB436 or E0771 cells pre-incubated (C) or not (NC) with adipocytes, treated (DOX) or not (NT) and then post-incubated (C) or not (NC) with adipocyte soluble factors. Right panel: quantification of the number of EVs secreted. d Analysis by flow cytometry of the DOX content in EVs secreted by MDA-MB436 or E0771 cells treated as in c. Abbreviation: a.u. arbitrary units.