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. 2004 Dec;78(24):13409–13419. doi: 10.1128/JVI.78.24.13409-13419.2004

FIG. 3.

FIG. 3.

Combination of Tyr mutation and cytoplasmic domain truncation enhances HIV Env protein stability on cell surfaces. Protein expression was carried out by transfection of HeLa cells. At 24 h posttransfection, the cells were pulse-labeled with [35S]Met-Cys labeling mix for 1 h and then chased in complete medium for 2, 4, or 6 h, followed by surface biotinylation and immunoprecipitation as described in Materials and Methods. Protein samples were prepared and analyzed by SDS-PAGE. (A) Pulse-chase radioactive labeling analysis for expression of HIV 89.6 Env and Env/Y712S. Lanes: 1, plasmid vector pCAGGS; 2, Env, pulse only; 3, Env, 2-h chase; 4, Env, 4-h chase; 5, Env, 6-h chase; 6, Env/Y710S, pulse only; 7, Env/Y710S, 2-h chase; 8, Env/Y710S, 4-h chase; 9, Env/Y710S, 6-h chase. (B) Pulse-chase radioactive labeling analysis for expression of HIV 89.6 Env750Tr and Env750Tr/Y710S. Lanes: 1, plasmid vector pCAGGS; 2, Env750Tr, pulse only; 3, Env750Tr, 2-h chase; 4, Env750Tr, 4-h chase; 5, Env750Tr, 6-h chase; 6, Env750Tr/Y710S, pulse only; 7, Env750Tr/Y710S, 2-h chase; 8, Env750Tr/Y710S, 4-h chase; 9, Env750Tr/Y710S, 6-h chase; Pre, HIV Env precursor gp160; SU, surface subunit gp120; TM, transmembrane subunit gp41.