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. 2020 Nov 6;10:19256. doi: 10.1038/s41598-020-75779-y

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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 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/.

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Effects of Tamoxifen treatment, cumulative (Model 1) and circulatory (Model 2) following co-incubation with MCF7 cells. MTAM Tamoxifen-treated MCF7 cells. MTAMWB Tamoxifen-treated MCF7 cells co-incubated with WB. MTAMPRP Tamoxifen-treated MCF7 cells co-incubated with PRP. MTAMPPP Tamoxifen-treated MCF7 cells co-incubated with PPP. WBTAMM Tamoxifen-treated WB co-incubated with MCF7 cells. PRPTAMM Tamoxifen-treated PRP co-incubated with MCF7 cells. PPPTAMM Tamoxifen-treated PPP co-incubated with MCF7 cells. (a) Platelet ultrastructural alterations. Black *—platelet hyalomere spread, white arrow—extending filipodia, black arrow—microparticles, black circle—fibrin clots and plaques with pores. A: MTAMWB—active platelets with a hyalomere spread and microparticles. B: WBMTAM—active platelets with a folded membrane and extending filipodia. C: MTAMPRP—platelets with a hyalomere spread and extending filipodia. D: PRPTAMM—platelets in early stages of aggregation, with extending filipodia. E: MTAMPPP—fibrin plaque formation with some pores. F: PPPTAMM—fibrin pores and remnants of fibrin. (b) Co-localisation of ERα (green) and ERβ (red) in MCF7 cells; following Tamoxifen-treatment and exposure to blood constituents (Model 1) and Tamoxifen-treatment of blood constituents exposed to MCF7 cells (Model 2). A: MTAM—cytoplasmic and nuclear ERα (arrow) and nuclear ERβ (*). B: MTAMWB—nuclear and cytoplasmic ERα (arrow) and nuclear ERβ (*). C: MTAMPRP—nuclear and low cytoplasmic ERα (arrow) and nuclear ERβ (*). D: MTAMPPP—low cytoplasmic and high nuclear ERα (arrow) and nuclear ERβ (*). E: WBTAMM—high cytoplasmic ERα (arrow) and extremely high nuclear ERβ (*). F: PRPTAMM—low cytoplasmic ERα (arrow) and high nuclear ERβ (white *), combined expression is noted (black *). G: PPPTAMM—cytoplasmic ERα with nuclear of ERβ (*). (c) Box and whisker plots representing quantitative analysis of ERα (green) and ERβ (red) expression in MCF7 cells for Model 1 and 2. ^[p < 0.05 between ERα and ERβ within the treatment group. ^#p < 0.05 between matched ERs compared to untreated MCF7. *p < 0.05 between matched ERs compared to Tamoxifen-treated MCF7. ^$p < 0.05 between matched ERs and blood constituent groups compared between models.