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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 the PKC/NOX downstream signaling pathway. (A) Washed platelets (3.6 × 10⁸ cells/ml) were pre-incubated with DMSO (solvent control), VAS compounds (2–10 μM), or Ro 31-8220 (2 μM) following stimulation with PDBu (150 nM) to trigger platelet aggregation. (B) The statistical analysis in (A). (C) After the reaction, platelet lysates were directly collected, and then subjected to Western blotting. Specific antibodies were used to detect PKC, IKKβ, and p38 MAPK. (D,E) Luciferase/luciferin and FITC-P-selectin antibody were used to detect ATP release and P-selectin using a microplate reader and flow cytometry, respectively. Data (B,D) are presented as means ± S.E.M. (n = 3). *p < 0.05, **p < 0.01, and ***p < 0.001, compared with the DMSO (solvent control) group. Data (C,E) are presented as means ± SEM (C, n = 4; E, n = 3). ***p < 0.001, compared with the resting group. ^##p < 0.01 and ^###p < 0.001, compared with the PDBu-treated (positive control) group. Comparisons were made by ANOVA.