CAP-PEs induce platelet aggregation and hyperreactivity in vitro and promote thrombosis in vivo in a TLR2-dependent manner. (A) Human platelets were isolated by gel filtration. A total of 2 × 10⁸/mL platelets were stimulated either with 1 U/mL thrombin, or 20 μM CAP-PEs (CEP-PE or CHP-PE) in the presence of 1 mg/mL fibrinogen and platelet aggregation was monitored in a Lumi-Aggregometer. (B-C) Platelet-rich plasma, isolated from WT (B) or TLR2^−/− (C) mice was primed with/without 20 μM CAP-PEs (CHP-PE), then stimulated with ADP and aggregation was monitored. (D) WT and TLR2^−/− mice were injected with 20 μM CAP-PEs (CEP-PE) or PE control via tail vein and after FeCl₃ injury; the time to complete thrombotic occlusion of carotid artery was recorded. (E) Presence of CAP-PEs (CEP-PE) in the plasma of ApoE^−/− mice fed chow diet (CD) or Western diet (WD) was determined by Far-Eastern blotting as described in the “Methods” section. (Right) Plasma concentrations of CEP-PE assessed by densitometry of Far-Eastern blotting using synthetic CEP-PE as a standard. (F) ApoE^−/−, ApoE^−/−/CD36^−/−, and ApoE^−/−/CD36^−/−/TLR2^−/− mice fed either CD or WD were used in intravital thrombosis assay. After FeCl₃ injury, the time to complete thrombotic occlusion of carotid artery was recorded. *P < .05, **P < .01, ***P < .001 and not significant, P > .05.