Transient expression of GPVI by transfected COS-7 cells. (A) The ligand blot assay was used to assess the binding of biotin-CVX to proteins in lysates of (from left to right): mock-transfected cells; cells transfected with wild-type (WT) GPVI; cells transfected with WT GPVI and then digested with the endoglycosidase PNGase F; and cells transfected with the substitution mutant N92A. The electrophoretic mobility of 2 representative marker proteins is indicated to the left of the gel. (B) The ligand blot assay used to assess the binding of biotin-CVX to proteins in lysates of (left) cells transfected with wild-type GPVI (–) and (right) cells transfected with wild-type GPVI then treated for 48 hours with tunicamycin (+). (C) Adhesion of COS-7 transfectants to CVX (▪) or CRP (□). The optical density (OD) is indicated on the x-axis. The identity of the transfectants is indicated on the ordinate and includes (from left to right): mock-transfected cells (COS-7); cells transfected with wild-type GPVI (WT); and cells transfected with N92A or S94A. Each bar represents the mean of 3 experiments. Error bars represent 1 SD.