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. 2002 Nov;76(22):11226–11235. doi: 10.1128/JVI.76.22.11226-11235.2002

FIG. 6.

FIG. 6.

Effect of Tsg101 expression on ubiquitination of HIV-2 Gag. (A) Schematic diagram of HIV-2 Gag and Tsg101 expression constructs. Abbreviations are as in Fig. 5. (B) 293T cells were transfected with constructs encoding a protease-negative, env-deleted HIV-2 provirus (pSVRGagΔNB) and either full-length Tsg101 (pTriExTsgΔHis), the N-terminal portion of Tsg101 (pTriEx5′TsgΔHis), the C-terminal portion of Tsg101 (pTriEx3′TsgΔHis), or control plasmid (pTriEx1.1ΔHis) in the presence or absence of a His-Myc-tagged ubiquitin expression construct (pH6M-Ub). At 48 h after transfection, protein extracts were applied to Ni-NTA columns, and ubiquitin-labeled HIV-2 Gag protein was eluted and detected by Western blotting with anti-p26 antibody. The positions of marker proteins are indicated in kilodaltons.