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

FIG. 7.

FIG. 7.

Cell surface expression of mutants containing six extra residues at the C terminus of the cytoplasmic tail. (A and B) Surface biotinylation of the 6His, 3(Leu-Ile), 3(Thr-Ser), and 6Gly mutants and of the 6His, 6Gln, 6Pro, and 6Trp mutants, respectively. HeLa cells transfected with the wild-type (WT) or mutant proviruses as indicated were metabolically labeled and surface biotinylated. Equal volumes of anti-HIV-1-isolated Env proteins were resolved by SDS-PAGE without (lanes 1 to 6) or with (lanes 7 to 12) prior precipitation with neutravidin-agarose. (C and D) FACS analyses of the 6His, 3(Leu-Ile), 3(Thr-Ser), and 6Gly mutants and of the 6His, 6Gln, 6Pro, and 6Trp mutants, respectively. VSV G-pseudotyped wild-type and mutant viruses as indicated were used to challenge CEM-SS cells, and total and cell surface Env expression was analyzed by FACS. Mock infection and infection with pseudotyped virus produced from pHXBCATΔBgl cotransfection, labeled Env−, were used as negative controls.