Effects of Cys mutations on formation of disulfide bonds between GP Ibα and GP Ibβ. (A) Illustration of wild-type and Cys mutant constructs of GP Ibα and GP Ibβ. (B-C) GP Ibα from the lysates of various transfected CHO cells that had been resolved in a 4-12% Bis-Tris SDS gel (Invitrogen) under nonreducing (N.R.) and reducing (R.) conditions. Molecular markers in kDa are labeled on the left. GP Ibα was blotted by WM23 antibody. (D) GP Ibα from various cell lysates that had been resolved in a 5% Tris-glycine SDS gel under nonreducing (N.R.) conditions. Dashed lines were added to show the difference between the protein bands. The figure is representative of 3 independent experiments.