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Figure 3 | Retrovirology

Figure 3

From: HIV-1 Group P is unable to antagonize human tetherin by Vpu, Env or Nef

Figure 3

Tetherin Counteraction by HIV-1 Group P Vpu and Nef. (A) FACS analysis of 293T cells cotransfected with a human or gorilla tetherin expression vector and pCGCG plasmids expressing GFP alone (GFP only) or together with the indicated vpu or nef alleles. (B) Reduction of Vpu- and Nef-mediated tetherin expression in 293T cells. Shown is the n-fold reduction of tetherin cell surface expression levels relative to those measured in cells transfected with the eGFP only control vector. The range of GFP expression used for the calculation is indicated in panel A. The mean (± SD) of two independent experiments is shown. (C) Western blot analysis of cell and virion lysates following cotransfection of 293T cells with a vpu- and nef-defective proviral HIV-1 NL4-3 construct, pCGCG plasmids expressing eGFP alone (eGFP only) or together with the indicated vpu or nef alleles and an empty vector (no tetherin) or human or gorilla tetherin expression plasmids. Cell and virion lysates were probed with an anti-HIV-1 capsid p24 monoclonal antibody. Sup., cell culture supernatant. (D) Infectious virus and (E) p24 release from 293T cells following cotransfection with a vpu- and nef-defective proviral HIV-1 NL4-3 construct, pCGCG plasmids expressing eGFP alone (eGFP only) or together with the indicated vpu or nef alleles and different amounts of human or gorilla tetherin expression plasmids. Infectious virus was determined by infection of TZM-bl reporter cells and p24 release was quantified by ELISA. Values are shown as percentage of values obtained in the absence of tetherin. All infections were performed in triplicates. The mean of two independent experiments is shown.

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