12-LOX–derived DPA_n-6oxylipins, 11-HpDPA_n-6and 14-HpDPA_n-6, do not impinge on the Gα_ssignaling pathway in platelets. Washed human platelets were treated with increasing concentrations of 11-HpDPA_n-6 (A) or 14-HpDPA_n-6 (B), along with DPA_n-6 (10 μM) or iloprost (1 μM) for 1 minute in the presence of IBMX (10 μM) (n = 2). (C) Human platelets (n = 5) were treated with dimethyl sulfoxide (DMSO), increasing concentrations of 11-HpDPA_n-6 or 14-HpDPA_n-6, DPA_n-6 (10 μM), 12(S)-HETrE (25 μM), or forskolin (1 μM) for 1 minute and immediately lysed, resolved on SDS-PAGE gel, and immunoblotted for VASP phosphorylation at S¹⁵⁷ or total VASP proteins. Data are means ± SEM. (D) Mouse platelets (n = 3) were treated with DMSO, 11-HpDPA_n-6 (1 μM), 14-HpDPA_n-6 (1 μM), DPA_n-6 (10 μM), 12(S)-HETrE (25 μM), forskolin (1 μM), or PAPA NONOate (5 μM) for 1 minute and immediately lysed, resolved on SDS-PAGE gel, and immunoblotted for VASP phosphorylation at S²³⁹ or total VASP proteins. Data are means ± SD. VASP phosphorylation levels were normalized to total VASP and DMSO control level to estimate the fold change.