12-HETrE is partially dependent on IP to inhibit human platelet activation. RO3244794- (250 nM) or vehicle control–treated human platelets were incubated with 12-HETrE (10 or 20 μM) or DMSO and then stimulated with U46619, the thromboxane mimetic (A), or collagen (B). One-way statistical analysis of variance with Dunn’s multiple comparison post-test was performed. Asterisks denote statistical differences between control- and RO3244794-treated groups: *P < .05, **P < .01, ***P < .001. Circles denote statistical differences between 12-HETrE–treated platelets and controls: •P < .05, ••P < .01, •••P < .001. (C) Human platelets were treated with DMSO, forskolin (5 µM), or 12-HETrE (10 or 25 µM) for 2.5 minutes and lysed, separated on a sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and immunoblotted with antibodies specific for phosphorylated (pS157; green) and total (red) VASP. The amount of phosphorylated VASP was normalized to the amount of total VASP in each sample, and data are reported as fold change in VASP phosphorylation relative to the DMSO treated sample. (D) VASP phosphorylation was measured by western blot using the lysates of human platelets incubated with RO3244794 or vehicle control and then treated with PGI₂ (4 nM), PGE₂ (20 μM), or PGD₂ (30 nM). (E) Human platelets pretreated with aspirin were stimulated with PAR4-AP (75 µM) in the presence of 12-HETrE (10 μM) or vehicle control. Data represent mean ± standard deviation. One-way statistical analysis of variance with Dunn’s multiple comparison post-test was performed. *P < .05, **P < .01, ***P < .001. ETOH, ethanol.