IP is required for 12-HETrE to inhibit mouse platelet activation. (A) Platelets isolated from WT (black bars) or IP^−/− (red bars) mice (n = 4-5) were stimulated with 25 or 50 µM of PAR4-AP in the presence of DMSO or 12-HETrE (10 or 25 μM). Two-tailed paired t test; ***P < .001. (B) The lysate of platelets from WT or IP^−/− mice that had been treated with DMSO, forskolin (Forsk; 5 µM), 12-HETE (HETE; 25 µM), or 12-HETrE for 2.5 minutes were separated on 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 DMSO-treated sample. One-way statistical analysis of variance with Dunn’s multiple comparison post-test was performed comparing DMSO with each condition. *P < .05, **P < .01, ***P < .001. Data represent mean ± standard deviation.