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. 2018 Oct 31;8:16094. doi: 10.1038/s41598-018-34409-4

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© The Author(s) 2018

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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Characterization of OVS isolated from oviductal fluids (OLF). (A) Protocol for isolation of oviductosomes (OVS) from OLF using ultracentrifugation. (B) Western blot reveals the presence of CD9 (24 kDa) on OVS from proestrus (Pro-OVS), metestrus (Met-OVS), and induced estrus (IE-OVS), as well as epididymosomes (EPS) used as a positive control. Each lane contains 40 μg of protein (n = 3). (C) Negative staining and TEM of OVS show the presence of membranous vesicles of both exosomal (<100 nm) and microsomal (100 nm–1 µm) sizes. (D) Immunogold labeling (6 nm gold particles) of CD9 is shown in OVS. Gold particles on individual OVS are seen arrowed on the exterior of the membrane (D-b-d). In the absence of primary antibodies and the presence of mouse IgG, gold particles were absent (D-a), indicating the specificity of the antibody. Scale bar = 100 nm.