Functional analysis of KI platelets. (A) Platelet aggregation study was assessed using platelet-rich plasma diluted to a concentration of 250 × 109/L. Data are representative of three independent experiments. (B) Activation state of αIIbβ3 and granular secretion were assessed by determining the JON/A binding (upper) and CD62P (lower) expression levels on platelets, respectively, with or without an agonist via flow cytometry. JON/A binding was adjusted by the expression level of CD41 and shown as %expression compared to that of WT. Data are presented as the mean and SD (n = 6 each). ∗∗∗P < .001 and ∗∗P < .01 (one-way ANOVA). (C) Washed platelets (2.5 × 105) from WT or Homo were added to the fibrinogen-coated coverslips with or without 20 μM ADP or 0.5 U/mL thrombin and incubated at 37°C for 20 min. Then platelets were fixed, permeabilized, and stained with TRITC-conjugated phalloidin. Scale bars represent 10μm. Representative results of 2 independent experiments are shown. (D) The number of adherent platelets (upper) and covered area (lower) were analyzed using the ImageJ software (National Institutes of Health). Results are presented as the mean and SD. ∗∗∗P < .001, ∗∗P < .01, and ∗P < .05 (unpaired t test).
