Figure 3.
Platelet aggregation and CLEC-2 tyrosine phosphorylation induced by cross-linking of CLEC-2 by anti–CLEC-2 antibody. (A) Human washed platelets (2 × 108/mL) were stimulated by 10 μ g/mL of anti–CLEC-2 antibody or control goat IgG in the presence of F(ab)′2 fragment of anti-FcγRIIA antibody (IV.3) and platelet aggregation was monitored with an aggregometer. (B) Human washed platelets (1 × 109/mL) were lysed with 2 × lysis buffer after stimulation as described. CLEC-2 was immunoprecipitated and sequentially Western blotted with antiphosphotyrosine antibody (4G10) or polyclonal anti–CLEC-2 antibody. The data are representative of 4 experiments. Lane 1: basal; lane 2: goat anti–CLEC-2; lane 3: control goat IgG.
