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. 2017 Jul 17;7:5582. doi: 10.1038/s41598-017-05817-9

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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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Post treatment of PGI₂ induces stress fibre reversal and actin nodule formation in platelets spread on fibrinogen in a time dependent manner. Platelets (2 × 10⁷/ml) were spread on 100 μg/ml fibrinogen for 25 minutes, washed with PBS, and then 10 nM of PGI₂ was added, for a further 2–60 minutes as per the representative experimental design in part (a) The platelets were then fixed and stained with FITC-phalloidin before being imaged via fluorescent microscopy. (b) Representative images of each condition of the experiment. (c) The number of spread platelets containing stress fibres was identified in control and PGI₂ treated samples. (d) The number of spread platelets containing actin nodules was identified in control and PGI₂ treated samples. (e) The average surface area of the spread platelets was analysed for each timepoint in control and PGI₂ treated samples. Analysis was performed using Image J. The experiments are an average of n = 5. p < 0.05. Scale-bar 5 μm. (f) Platelets were spread as above for 25 minutes and then treated for 10 minutes with either 10 nM PGI₂ or 1 μM forskolin. Assessment of the filamentous actin was performed using the F-actin assay. The experiments are an average of n = 3.