Figure 2
Figure 2. Role of PI3Kβ in functional platelet responses. (A) Role of PI3Kβ in promoting platelet aggregation. Platelets from p110flox/flox control (Cre−) or PF4-Cre/p110flox/flox (PF4-Cre+) mice were stimulated with thrombin, U46619, collagen, or convulxin, and aggregation was assessed using a Chrono-log dual-channel aggregometer under stirring at 900 rpm for 7 minutes. The profiles shown are representative of 5 independent experiments. Quantifications of the maximum of aggregation at 6 minutes are shown and are mean ± SEM of 5 independent experiments (right panels). (B) Role of PI3Kβ in promoting fibrin clot retraction. Photographs show the extent of clot retraction in wild-type and p110β-null platelet-rich plasma samples treated with 10 IU/mL thrombin. Quantification of the volume of serum extruded from the clot is shown. Results are the mean ± SEM of 3 independent experiments. (C) DIOC6-labeled platelets in whole blood from p110flox/flox control (Cre−) or PF4-Cre/p110flox/flox (PF4-Cre+) mice were perfused through fibrinogen-coated Bioflux plates at a shear rate of 1500 seconds for 5 minutes. Representative images at 5 minutes are shown. Images obtained with an epifluorescence microscope (Axiovert 200, Carl Zeiss; 40×/1.3 NA objective; 37°C) were captured with a CCD camera (CoolSnap HQ; Roper Scientific) and Metamorph software Version 6.2r6 (Universal Imaging Corp). Area covered by platelets was measured. Results shown are the mean ± SEM of 5 experiments. Statistical analysis: *P < .05; **P < .01; ***P < .005. (D) Thrombotic response of mice to ferric chloride injury of the carotid artery. Flow rates were measured in the carotid artery after exposure to 7% FeCl3 during 3 minutes. The experiment was stopped after 30 minutes. (i) For each genotype, the number of mice forming a stable occlusion is shown in black. The number of mice that formed an unstable occlusion that resolved is shown in gray. The number of mice that formed only a partial occlusion is shown in white. (ii) Representative flow traces for each case (stable occlusion, no occlusion, and unstable occlusion).

Role of PI3Kβ in functional platelet responses. (A) Role of PI3Kβ in promoting platelet aggregation. Platelets from p110flox/flox control (Cre−) or PF4-Cre/p110flox/flox (PF4-Cre+) mice were stimulated with thrombin, U46619, collagen, or convulxin, and aggregation was assessed using a Chrono-log dual-channel aggregometer under stirring at 900 rpm for 7 minutes. The profiles shown are representative of 5 independent experiments. Quantifications of the maximum of aggregation at 6 minutes are shown and are mean ± SEM of 5 independent experiments (right panels). (B) Role of PI3Kβ in promoting fibrin clot retraction. Photographs show the extent of clot retraction in wild-type and p110β-null platelet-rich plasma samples treated with 10 IU/mL thrombin. Quantification of the volume of serum extruded from the clot is shown. Results are the mean ± SEM of 3 independent experiments. (C) DIOC6-labeled platelets in whole blood from p110flox/flox control (Cre−) or PF4-Cre/p110flox/flox (PF4-Cre+) mice were perfused through fibrinogen-coated Bioflux plates at a shear rate of 1500 seconds for 5 minutes. Representative images at 5 minutes are shown. Images obtained with an epifluorescence microscope (Axiovert 200, Carl Zeiss; 40×/1.3 NA objective; 37°C) were captured with a CCD camera (CoolSnap HQ; Roper Scientific) and Metamorph software Version 6.2r6 (Universal Imaging Corp). Area covered by platelets was measured. Results shown are the mean ± SEM of 5 experiments. Statistical analysis: *P < .05; **P < .01; ***P < .005. (D) Thrombotic response of mice to ferric chloride injury of the carotid artery. Flow rates were measured in the carotid artery after exposure to 7% FeCl3 during 3 minutes. The experiment was stopped after 30 minutes. (i) For each genotype, the number of mice forming a stable occlusion is shown in black. The number of mice that formed an unstable occlusion that resolved is shown in gray. The number of mice that formed only a partial occlusion is shown in white. (ii) Representative flow traces for each case (stable occlusion, no occlusion, and unstable occlusion).

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