American Society of Hematology

Fig. 5.

$Fig. 5. VWF-stimulated complex formation between Src, PI 3–kinase, and GPIb is upstream of actin polymerization. / (A-B) Platelets were preincubated for 10 minutes at 37°C with 0.25% DMSO as control or 1 μM cytochalasin D (Cyto D), and stimulated with 6 μg/mL botrocetin plus 10 μg/mL VWF for the indicated time periods (seconds). The platelet lysates were immunoprecipitated with anti-GPIb (A) and anti-p85 (B) antibodies, respectively. The precipitated proteins were analyzed by anti-Src, anti-p85, and anti-GPIb immunoblotting. (C) Platelets without pretreatment with cytochalasin D were stimulated for 5 minutes and lysed. The lysates were then fractionated as described in Figure 3B. The Triton-insoluble fractions were further lysed in RIPA buffer, and the RIPA extracts were subjected to immunoprecipitation with anti-p85 antibody. The precipitated proteins were analyzed by anti-Src, anti-p85, and anti-GPIb immunoblotting.$

VWF-stimulated complex formation between Src, PI 3–kinase, and GPIb is upstream of actin polymerization.

(A-B) Platelets were preincubated for 10 minutes at 37°C with 0.25% DMSO as control or 1 μM cytochalasin D (Cyto D), and stimulated with 6 μg/mL botrocetin plus 10 μg/mL VWF for the indicated time periods (seconds). The platelet lysates were immunoprecipitated with anti-GPIb (A) and anti-p85 (B) antibodies, respectively. The precipitated proteins were analyzed by anti-Src, anti-p85, and anti-GPIb immunoblotting. (C) Platelets without pretreatment with cytochalasin D were stimulated for 5 minutes and lysed. The lysates were then fractionated as described in Figure 3B. The Triton-insoluble fractions were further lysed in RIPA buffer, and the RIPA extracts were subjected to immunoprecipitation with anti-p85 antibody. The precipitated proteins were analyzed by anti-Src, anti-p85, and anti-GPIb immunoblotting.

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