Characterization of ACC1^flx/flx, GPIbα-Cre^+/−, and ACC1 pKO mice. (A) Washed murine platelets or leukocytes were lysed and subjected to western blot analysis for ACC1 and ACC2 expression. Liver extract was used as positive control for ACC2. GAPDH served as loading control for platelet, leukocyte, and liver samples. (B-C) Platelet counts and MPV were determined by Cell-Dyn Emerald Hematology Analyzer (Abbott) in a single measurement conducted during the usage period of mice (8-16 weeks old). Data are expressed as mean ± standard error of the mean (SEM; n = ≥27). #P ≤ .0001 relative to ACC1^flx/flx mice, $P ≤ .0001 relative to GPIbα-Cre^+/− mice, and $§$P ≤ .0001 between ACC1^flx/flx and GPIbα-Cre^+/− mice. One-way analysis of variance (ANOVA) was used for data analysis. (D) Alexa 488–GPIbβ antibody was injected into ACC1^flx/flx, GPIbα-Cre^+/−, and ACC1 pKO mice to analyze platelet lifespan. Blood samples were collected on the day of the injection (day 0), as well as on days 3 and 5 after injection. Platelets were stained ex vivo with PE-conjugated anti-CD41 antibody, and the percentage of Alexa 488+ platelets among total CD41⁺ platelets was quantified by flow cytometry. Data are expressed as mean ± SEM (n ≥ 3). #P ≤ .05 relative to ACC1^flx/flx mice, $P ≤ .01 relative to GPIbα-Cre^+/− mice. Data were subjected to 2-way ANOVA analysis. (E) Platelet surface expression of αIIbβ3, GPIbα, and GPVI was measured by flow cytometry and normalized to MPV. Data are expressed as MFI adjusted to MPV (fL) ± SEM (n ≥ 8). #P ≤ .05 relative to ACC1^flx/flx mice, $P ≤ .01 relative to GPIbα-Cre^+/− mice, and $§$P ≤ .01 between ACC1^flx/flx and GPIbα-Cre^+/− mice. Data were subjected to 1-way ANOVA analysis. GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GPVI, glycoprotein VI; MFI, median fluorescence intensity; MPV, mean platelet volume.