Fig. 2.
Association of 14-3-3 with GP Ib-IX complex.
One milliliter cytosol or the RIPA-extracted actin cytoskeletal (actin CSK) fraction from resting, thrombin-activated (1 U/mL) (A), or vWF- and botrocetin-activated platelets (B) was immunoprecipitated using 5 μL 14-3-3ζ antibody or nonimmune sera (Non I) (5 μL) and was immunoblotted using antibodies to the extracellular domain of GP Ib (anti-glycocalicin). As indicated, in some experiments the platelet subcellular fractions were preincubated with 1 mmol/LN-ethylmaleimide (NEM) to displace actin-binding protein. This figure is representative of 5 similar experiments.