Figure 3.

Lec1 cells secreted ADAMTS13 proteins with oligomannose N-glycans. ADAMTS13 (WT) and its truncated variants, AD5 and AD2, were expressed in CHO or the Lec1 mutant cells (Lec1). Cell lysates (C) and serum-free culture media (M) were collected for SDS-PAGE and immunoblotting for V5 tag 72 hours after transfection. Each secreted protein was also digested with PNGase F (F) or endoglycosidase H (H) before analysis. The numbers to the left of each gel indicate the position of the molecular weight markers in kilodaltons. Smaller molecular weight bands in the cell lysates of WT and AD2 might have resulted from proteolysis of expressed protein.