Treatment of wild-type GPVI with endoglycosidases or tunicamycin. (A) A shift in the electrophoretic mobility of wild-type GPVI was observed following treatment with endoglycosidases. MAb LJ6.5 was used in a Western blot to visualize GPVI in lysates of (from left to right): cells transfected with wild-type (WT) GPVI; cells transfected with WT GPVI and then digested with the endoglycosidase Endo H, which cleaves high mannose type N-glycans; cells transfected with WT GPVI and then digested with the endoglycosidase PNGaseF, which cleaves complex type N-glycans; 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 influence of tunicamycin treatment on Dami cell adhesion. The adhesion of Dami lines transfected with vector alone (mock) or WT GPVI was compared to that of same Dami lines transfected with WT GPVI but first exposed to tunicamycin for 48 hours. Adhesion to plates coated with BSA (▦), CVX (▪) or CRP (□) was measured. Optical density (OD) is indicated on the abscissa. Error bars represent 1 SD.