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. 2019 Dec 11;9:18852. doi: 10.1038/s41598-019-55189-5

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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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Effects of VAS compounds on calcium mobilization and GPIIbIIIa activation. (A,B) Washed platelets were pre-incubated with DMSO (solvent control) and VAS1 (2–10 μM) before the addition of PDBu to trigger platelet activation. Fura-2 and FITC-PAC1 antibodies were used to detect calcium mobilization and GPIIbIIIa activation by a microplate reader and flow cytometry, respectively. (C) FITC-triflavin was used to observe the binding capacity of VAS compounds (10 μM) on the GPIIbIIIa receptor through flow cytometry. (D) LDH assay kits were used to determine the cytotoxicity of VAS compounds (10–100 μM) on platelets. The Triton-treated group as maximum toxicity (100%). Data (A) are presented as means ± SEM (n = 3). *p < 0.05 and ***p < 0.001, compared with the DMSO (solvent control) group. Data (B) are presented as means ± S.E.M. (n = 3). ***p < 0.001, compared with the resting group. ^#p < 0.05 and ^##p < 0.01, compared with the PDBu-treated (positive control) group. Comparisons were made by ANOVA. Profiles (C) are representative examples of three similar experiments.