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. 2021 Jul 28;11:15308. doi: 10.1038/s41598-021-94052-4

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Inhibiting sphingosine kinases reduces PAR1-AP induced platelet aggregation. Washed human platelets (2 × 10⁸/mL) were pre-incubated with vehicle control (0.2% methanol) or sphingosine kinase inhibitors for 5 min. Platelet aggregation was induced by maximal (1.5–2.0 μM) concentration of PAR1-AP and recorded for 5 min. (a) Representative traces of maximal concentration PAR1-AP induced platelet aggregation in the absence or presence of (i) 10 μM SPHK1/2 inhibitor DMS (N, N-Dimethylsphingosine), (ii) 1 μM SPHK1 inhibitor PF543 hydrochloride or (iii) 10 μM SPHK2 inhibitor SLM 6,031,434, respectively. (b) Bar graphs of (i) quantified percentage aggregation and (ii) AUCs induced by maximal concentration of PAR1-AP alone or in the presence of SPHK inhibitors. AU: arbitrary unit. Data are plotted as mean ± standard error of the mean. N = 5. Statistical analysis: Student’s paired t-test. ns = not significant, *P < 0.05, **P < 0.01, ***P < 0.001.