American Society of Hematology

Figure 1

$Figure 1. CLEC-2 translocates to lipid rafts upon stimulation. (A) Resting and rhodocytin-stimulated platelets were lysed in the presence of 1% Brij 58 and separated into lipid raft and nonraft fractions by sucrose gradient ultracentrifugation. The localization of CLEC-2 was determined by immunoblotting with specific antibodies. The lipid raft fractions were identified using an anti-LAT blot, and nonraft fractions were identified using an anti–integrin αIIb blot. (B) Characterization of fractions from basal and rhodocytin stimulated platelets pretreated with DMSO or 10μM cytochalasin D. Data represent the mean and standard error of 3 independent experiments. (C) CLEC-2 was immunoprecipitated from lipid raft and nonraft fractions of platelets stimulated with 100nM rhodocytin and blotted with an antiphosphotyrosine antibody. Membranes were subsequently stripped and reblotted with anti–CLEC-2. Data are representative of at least 3 independent experiments.$

CLEC-2 translocates to lipid rafts upon stimulation. (A) Resting and rhodocytin-stimulated platelets were lysed in the presence of 1% Brij 58 and separated into lipid raft and nonraft fractions by sucrose gradient ultracentrifugation. The localization of CLEC-2 was determined by immunoblotting with specific antibodies. The lipid raft fractions were identified using an anti-LAT blot, and nonraft fractions were identified using an anti–integrin α_IIb blot. (B) Characterization of fractions from basal and rhodocytin stimulated platelets pretreated with DMSO or 10μM cytochalasin D. Data represent the mean and standard error of 3 independent experiments. (C) CLEC-2 was immunoprecipitated from lipid raft and nonraft fractions of platelets stimulated with 100nM rhodocytin and blotted with an antiphosphotyrosine antibody. Membranes were subsequently stripped and reblotted with anti–CLEC-2. Data are representative of at least 3 independent experiments.

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