Figure 4
From: A suicide gene approach using the human pro-apoptotic protein tBid inhibits HIV-1 replication
Inhibition of HIV-1 NL4-3 replication in pLtBid(INS) 2 R-CD4 transfected HeLa SS6 cells. HeLa SS6 cells were transfected with the control vector pLRed(INS)2R-CD4 or the suicide vector pLtBid(INS)2R-CD4. 36 hours after transfection, HeLa SS6 cells were infected with HIV-1 NL4-3. The data shown are from two independent experiments, each performed in duplicate. (A) CD4 expression was determined by FACS analysis 24 hours after infection with HIV-1 NL4-3 in mock transfected (dotted line), pLRed(INS)2R-CD4 transfected (black line), and pLtBid(INS)2R-CD4 transfected (grey line) HeLa SS6 cells. (B) Release of viral particles was determined by p24-ELISA using cell-free supernatants of HIV-1 NL4-3 infected HeLa SS6 cells transfected with pLRed(INS)2R-CD4 (open triangles), pLtBid(INS)2R-CD4 (black circles), or mock controls (open circles). Shown is one representative assessment. (C) Depicted is the residual replication of HIV-1 NL4-3 at days 3 and 4 post infection in pLtBid(INS)2R-CD4 transfected HeLa SS6 cells (black) as measured by p24 antigen ELISA, background subtracted, and normalized to p24 values obtained in pLRed(INS)2R-CD4 transfected HeLa SS6 cells (white). (D) Determination of cell death in HeLa SS6 cells in mock transfected (grey), pLRed(INS)2R-CD4 transfected (white), and pLtBid(INS)2R-CD4 transfected (black bar) cells 48 h after infection with HIV-1 NL4-3. Percentages of DCD-stained cells are depicted.