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. 2010 Feb 24;84(9):4851–4855. doi: 10.1128/JVI.01699-09

FIG. 1.

FIG. 1.

CsA disrupts HIV-1 Env glycoprotein incorporation into virions if the drug is present during virion production. HIV-1 virions were produced in 293T cells by transfection of pNL4-3 (A), with pNL4-3-env− and one of the indicated Env proteins (B), or with pNL4-3-env− and either HIV-1 Env (C and F), VSV G (D and G), or ampho-MuLV Env (E and H). Drugs were added to the medium during virion production. For panel A, CsA was added at the indicated concentrations. For panels B to H, the indicated drugs were added at 10 μM CsA. Virion-containing supernatant was filtered, and virions were enriched by ultracentrifugation through a 25% sucrose cushion. Virion preparations were normalized by reverse transcription and tested for infectivity (A and B) or subjected to Western blotting using antibodies against the indicated proteins (C to E). At 48 h posttransfection, the producer cells were harvested using 5 mM EDTA, normalized by cell number, and subjected to Western blotting using antibodies against the indicated proteins (F to H).