PKM2 regulates PI3K-mediated Akt/GSK3β signaling in platelets. (A) Human and mouse platelets were pretreated with vehicle or ML265 (50 and 100 μM) for 10 minutes at room temperature before stimulation with convulxin (100 ng/mL) or thrombin (0.1 U/ml) for 10 minutes. The left panels show representative western blots for phospho-Akt (Ser 473), total Akt, phospho-GSK3β, and total GSK3β. The middle and right panels show densitometry analysis of immunoblots. Values are mean ± SEM, n = 4 per group. *P < .05 vs resting platelets, ^†P < .05 vs activated platelets (vehicle). Two-way ANOVA with Tukey’s multiple comparisons test. (B) Platelets from PKM2^fl/fl or PKM2^fl/flPF4Cre⁺ were stimulated with convulxin (100 ng/mL) or thrombin (0.1 U/mL) for 10 minutes. The left panels show representative western blots for phospho-Akt (Ser 473), total Akt, phospho-GSK3β, and total GSK3β; the middle and right panels show densitometry analysis of immunoblots. Values are mean ± SEM, n = 3 to 4 per group. *P < .05 vs resting platelets, ^†P < .05 vs PKM2^fl/fl. Two-way ANOVA with Tukey’s multiple comparisons test. RP, resting platelets.