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. 2006 Aug;80(16):8047–8059. doi: 10.1128/JVI.00252-06

FIG. 4.

FIG. 4.

Suppression of CD4 expression in transfected 293T cells. (A) 293T cells were cotransfected with the indicated HIV-1 IRES-eGFP proviral constructs and a CD4 expression construct and analyzed by flow cytometric analysis 2 days later. Numbers are the rMFIs at the indicated ranges of eGFP expression. (B) Average suppression of CD4 expression (± SD, n = 3) by expression of the indicated HIV-1 proviral genomes. The rMFI obtained for the vpu-, env-, and nef-defective construct (panel A, lane 1) was divided by the rMFI obtained for the remaining proviral constructs to calculate suppression (n-fold) of CD4 surface expression. The results were confirmed in two independent experiments. (C) Correlation between down-modulation of CD4 in HIV-1-infected Jurkat T cells (measured at 48 h posttransduction at medium levels of eGFP expression) (Fig. 2A) and suppression of CD4 expression by transfected 293T cells. (D) Inhibitory effects of Vpu, Env, and Nef at different levels of CD4 expression. 293T cells were cotransfected with 4 μg of the proviral constructs and 0.1, 0.5, and 1.0 μg of the CD4 expression plasmids. E, env; U, vpu; N, nef; uninf., uninfected; Mock, mock infected; suppr., suppressed; down-mod., down-modulated.