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. 2020 Oct 6;22:105. doi: 10.1186/s13058-020-01342-2

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© The Author(s) 2020

Open AccessThis 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. 2 — Trastuzumab upregulates PERP in extracellular vesicles emitted by trastuzumab-sensitive but not by trastuzumab-resistant human breast tumor cells. BT-474 (a) or BT-474TR (b) cells were treated (+) or not (−) with 5 μg/ml trastuzumab (TZ) in 3D culture for 72 h. Extracellular vesicles were isolated from the cell media by ultracentrifugation and assayed for PERP levels by western blot. An extracellular vesicle marker Flotillin-1 is a loading control. Trastuzumab-sensitive MCF7/Her2-18 (c) and trastuzumab-resistant HCC-1419 (d) human ErbB2-positive breast cancer cells were treated (+) or not (−) with trastuzumab for 48 h (c) or 72 h (d), and extracellular vesicles were isolated from the cell media as in a and analyzed as in a. Bar graphs to the right of the western blot images represent quantification of respective bands. PERP levels were normalized by the levels of the loading control. PERP levels (relative level) in the control cells were designated as 1.0. The data represent the average of five (a), three (b, c), and two (d) independent experiments, plus SE. *p value was < 0.05