FIGURE 2.

Glycosylation of Wise. A–D, conditioned media from cells transfected with Wise (A), Wise(ΔC) (B), Wise(ΔN) (C), or Wise(N47A) (D), treated with O-glycosidase or PNGase F. Wise and Wise(ΔC) are sensitive to PNGase F and not to O-glycosidase. In B, the cell extract was also loaded, showing that the majority of Wise protein in the cell extract is a non-glycosylated form, and only a small amount of the glycosylated form exists. Wise(ΔN) and Wise(N47A) are not sensitive to PNGase F. E, conditioned medium of Wise treated with endo-β-N-acetylglucosaminidase D (Endo D) or endo-β-N-acetylglucosaminidase H (Endo H). Wise is sensitive to endo-β-N-acetylglucosaminidase D and resistant to endo-β-N-acetylglucosaminidase H.