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Fig. 7. The integrity of lipid rafts is required for efficient platelet fibrin clot retraction. (A) Control showing raft disruption by cholesterol depletion (left panel). The level of LAT and CD36 in raft (pooled fractions 2-4 of the sucrose gradient) isolated from platelet pre-treated or not with 5 mM MßCD for 10 minutes (leading to 50±2% depletion of total cholesterol and 56±4% depletion of cholesterol in rafts) was analyzed by western blotting. The effect of raft disruption on platelet aggregation induced by low (0.2 IU/µl) or high (0.5 IU/µl) thrombin concentration is shown in the right panel. (B) Effect of raft disruption on fibrin clot retraction. Platelets treated or not with 5 mM MßCD for 10 minutes, were resuspended at 3x10⁸/ml in 2 ml autologous plasma prior to treatment with either 0.5 IU/ml thrombin and 0.1 µg/ml atroxin or 2 IU/ml thrombin alone at 37°C. For the study with 0.5 IU/ml thrombin, atroxin was added to induce clot formation. After 2 hours, the extent of clot retraction was observed.

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