American Society of Hematology

Fig. 1.

$Fig. 1. Activation-dependent translocation of PI 3-kinase, GP Ib, and 14-3-3 to the cytoskeleton. / Washed platelets were stimulated with thrombin (1 U/mL) (A) or vWF and botrocetin (C) for the indicated times. Triton-soluble cytosol (Cytosol), actin cytoskeletal (actin CSK), and membrane cytoskeletal (membrane CSK) fractions were isolated. Thirty microliters of each fraction was analyzed by SDS-PAGE and immunoblot analysis using antibodies to glycocalicin (GP Ib), the p85 subunit of PI 3-kinase (p85), or 14-3-3ζ. (B) Platelets were stimulated with thrombin (1 U/mL), with or without stirring, or were preincubated with 2 mmol/L RGDS for 10 minutes before thrombin stimulation for 0, 3, or 5 minutes. Thirty microliters of the cytosolic or actin cytoskeletal fractions was analyzed by SDS-PAGE and by immunoblot analysis using antibodies to p85 or 14-3-3ζ. This figure is representative of 3 similar experiments.$

Activation-dependent translocation of PI 3-kinase, GP Ib, and 14-3-3 to the cytoskeleton.

Washed platelets were stimulated with thrombin (1 U/mL) (A) or vWF and botrocetin (C) for the indicated times. Triton-soluble cytosol (Cytosol), actin cytoskeletal (actin CSK), and membrane cytoskeletal (membrane CSK) fractions were isolated. Thirty microliters of each fraction was analyzed by SDS-PAGE and immunoblot analysis using antibodies to glycocalicin (GP Ib), the p85 subunit of PI 3-kinase (p85), or 14-3-3ζ. (B) Platelets were stimulated with thrombin (1 U/mL), with or without stirring, or were preincubated with 2 mmol/L RGDS for 10 minutes before thrombin stimulation for 0, 3, or 5 minutes. Thirty microliters of the cytosolic or actin cytoskeletal fractions was analyzed by SDS-PAGE and by immunoblot analysis using antibodies to p85 or 14-3-3ζ. This figure is representative of 3 similar experiments.

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