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. 2004 May;78(10):5157–5169. doi: 10.1128/JVI.78.10.5157-5169.2004

FIG. 6.

FIG. 6.

Analyses of N-linked glycosylation of the 6His mutant. (A) PNGase F analysis. Equal volumes of MAb 902-isolated Env proteins from HeLa cells transfected with the wild-type (WT) or 6His mutant provirus were treated or not with PNGase F prior to Western blotting with MAb 902. Bands a and b represent the gp160 and gp120 forms without N-glycans, respectively. (B) DMM treatment. Cells transfected with the wild-type or mutant provirus were grown in the presence or absence of 1 mM DMM for 48 h, and cell lysates were analyzed by Western blotting with MAb 902. Bands a and b represent the high-mannose forms of gp120 and gp41, respectively. (C) Neuraminidase digestion. Equal volumes of the isolated wild-type and 6His mutant Env proteins were treated or not with neuraminidase prior to Western blotting with MAb Chessie 8. (D) Pulse-chase and endo H digestion. HeLa cells transfected with the wild-type or 6His mutant provirus were metabolically labeled and chased for different times. Equal volumes of MAb 902-isolated Env proteins were treated or not with endo H prior to Western blotting with MAb 902.