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. 2017 Aug 16;7:8476. doi: 10.1038/s41598-017-09158-5

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© The Author(s) 2017

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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Localization of myosin IC isoform A in prostate cancer cells and exosomes. (A) Western blot analysis of total cell (TC) extract and secreted exosomes (Ex) of PC3 cells. Left labels: molecular weight in kDa. Right labels: antibody targets. (B) Western blot analysis of total extract and exosomes of PC3 cells transfected with EGFP-tagged isoforms A and B of myosin IC or with EGFP alone. Top panels: probed with anti-EGFP antibody. Bottom panels: probed with anti-myosin IC (non-isoform specific) antibody. (C) Fluorescence microscopy images of purified exosomes of PC3 cells transfected with EGFP-tagged isoform A or EGFP alone. Membrane visualized with Vybrant DiI dye. Arrows: vesicles displaying colocalization of the two fluorescent signals. Zoom: central area of image to the left, magnified. (D) Relative localization of EGFP-tagged isoform A with caveolin 1 in PC3 cells. Caveolin is visualized by indirect immunofluorescence. WT, wild type. K892A, substitution in the lipid-binding domain. Scale bars in (C,D) – 10 µm.