Fig. 3.
Inhibition of thrombin-induced BMS-650032 stress fiber formation and platelet spreading by incorporation of anti-mDia1 antibody. Untreated platelets (control), platelets treated with DMSO alone (DMSO), platelets incorporated with a control IgG (cIgG), and platelets loaded with anti-mDia1 antibody were all allowed to spread on fibrinogen-coated coverslips in the presence of thrombin (1 IU/ml) for 45 min. After fixation and permeabilization, platelets were stained for F-actin and mDia1 (A) or for F-actin only (B). Original magnification, 1000×. The cell surfaces (C) were quantified as described in Materials and methods. Data are expressed as the means ± SE of at least three independent experiments. ?P platelets were harvested by scraping into lysis buffer. GTP–RhoA (A,B) and its binding to mDia1 (D,E) were determined in cell lysates subjected to immunoprecipitation and immunoblotting analysis as described in Materials and methods. The cell surfaces (C) were quantified as described in Materials and methods. Data are expressed as the means ± SE of at least three independent experiments. ?P
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Immunohistochemistry and assessment Antibodies used were as follows IL goat
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