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. 2020 Aug 7;10:13360. doi: 10.1038/s41598-020-70302-9

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© The Author(s) 2020

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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Western blot analysis and relative quantification of GM-CSF expression in porcine genital tract, epididymal fluid (EF), seminal plasma (SP) and in spermatozoa from cauda epididymis (CE) and different ejaculate portions. In (a1), three bands of GM-CSF expression with the different glycosylation degrees: 15, 31 and 40 kDa. In (a2–a4), the relative quantity of 15 kDa (a2), 31 kDa (a3) and 40 kDa (a4) in reproductive tissues; T: testes. CaE: caput epididymis. CoE: corpus epididymis. CauE: cauda epididymis. P: prostate. SV: seminal vesicle. B: bulbourethral gland. In (b1) one band of active GM-CSF expression of 15 kDa. In (b2) relative quantity of 15 kDa in EF and SP from the different ejaculate portions: first 10 mL-SRF, rest of SRF, post-SRF, entire ejaculate (EE). (c1) Two bands of active and glycosylated GM-CSF of 15 and 31 kDa. (c2 and c3) Relative quantity of 15 kDa (c2) and 31 kDa (c3) in mature spermatozoa from CE, first 10 mL-SRF, rest of SRF, post-SRF and EE. L: expression in liver tissue, as control. ^a–d Indicate relative quantitative differences (p < 0.05) among tissues, fluid and sperm sources. SRF: sperm-rich fraction. Cropped image of Western blot (see Supplemental Information, Fig S6 for full image).