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. 2014 Feb 14;289(13):9146–9157. doi: 10.1074/jbc.M113.534321

FIGURE 4.

FIGURE 4.

HIV-2 Vpx inhibits IRF5 transactivation activity. A, 293T cells in a 12-well plate were transfected with pRL-TK control plasmid (100 ng), IL12p40-luc reporter plasmid (300 ng), and IRF5 expression plasmid (1 μg) together with increasing amounts of Vpx expression plasmid (0, 1, and 3 μg) as indicated. Immunoblot analysis of whole cell lysates prepared for luciferase assay is shown below. B, HT1080 cells in a 12-well plate were transfected with pRL-TK control plasmid (200 ng), IL12p40-luc reporter plasmid (600 ng), and IRF5-expressing plasmid (1.5 μg) together with increasing amounts of Vpx expression plasmid (0, 1.5, and 4.5 μg) as indicated. Immunoblot analysis of whole cell lysates prepared for luciferase assay is shown below. C, 293T cells in a 12-well plate were transfected with pRL-TK control plasmid (100 ng), IL12p40-luc reporter plasmid (300 ng), IRF5 (1 μg), and MyD88 (0.5 μg) expression plasmids together with increasing amounts of Vpx expression plasmid (0, 2, and 6 μg) as indicated. Immunoblot analysis of whole cell lysates prepared for luciferase assay is shown below. In all transfections, the pCNF vector was added to bring the total plasmid to the same amount. Luciferase activity was measured at 24 h post-transfection by Dual-Luciferase reporter assay. Relative luciferase activity was determined as fold induction (relative to the basal level of reporter genes in the presence of pCNF vector after normalization with co-transfected relative luciferase units). Values are mean ± S.D. for three experiments.