FIGURE 5.

gp120 glycosylation is influenced by metabolic manipulations. Jurkat and HepG2 cells stably transfected with gp120-encoding DNA plasmid were cultured in the presence of N-acetylglucosamine, 80 mm (GlcNAc); uridine, 5 mm (Uri); N-acetylglucosamine 80 mm plus uridine 5 mm (Uri+GlcNAc); succinate, 20 mm (S), pyruvate, 4.5 mm (Pyr); Gal, 50 mm; Man, 50 mm; DMJM, 800 μm; or mock-treated (Untr, negative control). gp120 was purified using Ni-NTA-agarose, separated by 10% SDS-PAGE, blotted, and developed with anti-V5-tag antibody (A). Purified gp120 glycoproteins were treated with PNGase F (*), Endo H (**), or mock treated (***), and the resultant preparations were analyzed by SDS-PAGE and Western blotting with anti-V5-tag antibody. gp120 was from Jurkat (B) and HepG2 cells (C). Representative results from two experiments are shown.