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. 2018 Jul 9;8:10300. doi: 10.1038/s41598-018-28211-5

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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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MC reveals novel alterations in the platelet surface expression of antigens in Glanzmann thrombasthenia (GT) patients. Citrate-anticoagulated blood samples from healthy donors (n = 3) and a GT patient (3 separate blood draws from the same patient on 3 different visits) were treated with vehicle, ADP (0.5 or 20 µM) or TRAP (1.5 or 20 µM) for 30 minutes in the presence of a fluorescent-tagged antibody cocktail (A) (CD41a-PE and CD61-FITC or PAC1-FITC and CD62P-PE) or a custom metal-tagged antibody cocktail (B) (CD41-149Sm, CD61-165Ho, PAC1-159Tb, CD62P-172Yb, CD63-161Dy, CD9-171Yb, CD154-154Sm, CD42a-155Gd, CD42b-163Dy, GPVI-152Sm, CD31-145Nd, CD36-150Nd, CD29-176Yb and CD107a-166Er). Samples were fixed in 1% formaldehyde and analyzed by flow cytometry or mass cytometry. Results are expressed as a percentage of the mean fluorescence intensity (MFI; flow cytometry readout) or mean metal intensity (MMI; mass cytometry readout) achieved with 20 µM TRAP in healthy donor platelets (means ± SEM; n = 3). Statistical analysis: 2-way ANOVA was used in conjunction with a Bonferroni post-test to indicate statistical significance; *P < 0.05, **P < 0.01 and ***P < 0.001. Abbreviations: ADP, adenosine diphosphate; FITC, fluorescein isothiocyanate; MC, mass cytometry; PE, phycoerythrin; TRAP, thrombin receptor activating peptide.