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. 2019 Dec 12;9:19009. doi: 10.1038/s41598-019-55391-5

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© The Author(s) 2019

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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Representative 2D-DIGE gels pH 4–7 and pH 6–9 with significantly altered platelet proteins upon inhibition and activation are delineated. These exemplary monochrome-displayed 2D-DIGE images show significantly altered platelet protein spots upon inhibition and activation which are discussed in detail in the manuscript. In Supplementary Fig. S2 coloured 2D-DIGE images are displayed. A total of 36 µg (12 µg sample Cy3-, 12 µg sample Cy5-, and 12 µg internal standard Cy2-labeled) platelet protein extracts were separated according to the isoelectric point (pI) and the molecular weight (MW) in the pH ranges (A) pH 4–7 and (B) pH 6–9. Altered proteins were identified by mass spectrometry and are outlined for inhibition and for activation with their respective UniProt “Gene” name. Protein spots of interest were selected according to (a) protein spots matched >90% of all 2D-DIGE gels, (b) protein abundance changes >20% between treatment group and untreated control and (c) FDR-corrected ANOVA p-value <0.05.