ACC1 pKO platelets displayed reduced TxA₂ generation, dense granule secretion, and aggregation upon agonist stimulation. (A-G) Washed murine platelets from ACC1 floxed or pKO mice were stimulated with various Thr or CRP concentrations. (A) Platelets were stimulated with 100 mU/mL Thr or 0.3 μg/mL CRP for 5 minutes, and TxB₂ was measured in the supernatant using an enzyme-linked immunosorbent assay kit. Aspirin-treated platelets were included as a positive control for TxB₂ generation inhibition in the CRP condition. Data are expressed as mean ± SEM (n = 4). ∗P ≤ .05 relative to unstimulated conditions. #P ≤ .05 relative to ACC1^flx/flx mice. Data were analyzed via 2-way ANOVA. (B) Aggregation was analyzed by turbidimetry (Chrono-Log). Data are expressed as mean ± SEM (n ≥ 4). ∗P ≤ .05 relative to the first agonist concentration. #P ≤ .05 relative to ACC1^flx/flx mice. Two-way ANOVA was used for analysis. (C) Aggregation profiles are shown. (D) Activated αIIbβ3 (JON/A) and (E) p-selectin (CD62P) exposure was detected by flow cytometry. Data are expressed as fold change ± SEM (n = 4) and mean ± SEM (n = 4) respectively. ∗P ≤ .05 relative to unstimulated conditions. #P ≤ .05 relative to ACC1^flx/flx mice. Two-way ANOVA was used for analysis. (F) Dense granule secretion was assessed via addition of luciferase-luciferin reagent (Chrono-log). Data are expressed as mean ± SEM (n ≥ 3). ∗P ≤ .05 relative to the first agonist concentration. #P ≤ .05 relative to ACC1^flx/flx mice. Two-way ANOVA was used for analysis. (G) Profiles are shown. ASA, aspirine; Thr, thrombine.