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Figure 1


Fig. 1. Subcellular localization of DAB2 in human platelets. (A) Association of DAB2 with platelet {alpha}-granules. Platelet homogenates (2x109) were separated by sucrose-density-gradient (30-60%) centrifugation. Eighteen fractions (700 µl each) were collected from the top and aliquots were subjected to western blot analysis using the anti-p96 (DAB2), anti-P-selectin and anti-PF4 antibodies. (B) Colocalization of DAB2 with PF4. Immunofluorescence staining of DAB2 (green) and PF4 (red) was performed with platelets cytospun on a glass slide and was observed by confocal microscopy. The image of individual platelets was obtained by using confocal microscopy analytical software to enlarge and focus the indicated platelet. (C) Immunoelectron microscopy of resting platelets revealed the presence of DAB2 in {alpha}-granules. Washed platelets were immunostained with anti-DAB2 antibody. After labeling with 10-nm colloidal-gold-conjuagated protein G, the subcellular distribution of DAB2 was observed by electron microscopy. Arrows indicate positive staining of the immunogold particle. Original magnification 15,000x.





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