DPA_n-6and its oxylipins, 11-HpDPA_n-6and 14-HpDPA_n-6, impair in vivo thrombus formation in laser-induced cremaster arteriole thrombosis models. (A) Representative images of platelet accumulation (green) and fibrin formation (red) in growing thrombi in cremaster arterioles in WT control treated with vehicle control (ctrl), DPA_n-6, 11-HpDPA_n-6, or 14-HpDPA_n-6. Time after vascular injury is indicated above (n = 3 mice, 10-12 thrombi per mouse). Scale bars, 10 μm. (B) Dynamics of platelet accumulation in thrombi analyzed by change in fluorescent intensity following injury. (C) Dynamics of fibrin formation within thrombi analyzed by change in fluorescent intensity following injury. (D) Representative images of platelet accumulation (green) and fibrin formation (red) in growing thrombi in cremaster arterioles in PPARα^−/− mice treated with vehicle control (ctrl), DPA_n-6, 11-HpDPA_n-6, or 14-HpDPA_n-6. Time after vascular injury is indicated above (n = 3 mice, 10-12 thrombi per mouse). (E) Dynamics of platelet accumulation in thrombi in PPARα^−/− mice analyzed by change in fluorescent intensity. (F) Dynamics of fibrin formation in thrombi in PPARα^−/− mice analyzed by change in fluorescent intensity following injury. (G) Mean tail bleeding of control mice (n = 24) or mice treated with DPA_n-6 (n = 17) is denoted by the horizontal line. (H) The proposed model of DPA_n-6 12-LOX oxylipins’ inhibitory effect on platelet activation. DPA_n-6 12-LOX oxylipins derived from the elongation of DGLA into the bioactive lipids 11-HpDPA_n-6 and 14-HpDPA_n-6. DPA_n-6 oxylipins regulate platelet function through activation of PPARα, which leads to inhibition of dense and α-granule release, calcium mobilization, and platelet reactivity in response to G protein–coupled receptor–based or immunoreceptor tyrosine-based activation motif–mediated platelet activation. Data are means ± SEM. ****P < .0001, 2-way ANOVA.