Figure 4

Vpu suppresses HIV-1 transcription. A. 293T cells were transfected with pNL4-3 and increasing amounts of Vpu expression plasmid. After 48 hours a luciferase assay was performed to determine the effect of Vpu of HIV-1 transcription. Data are normalized to empty vector control (EV). B. Transfected 293T cells were harvested, lysed and equal amounts of cell lysate protein were subjected to 4-12% SDS-PAGE. After transfer to nitrocellulose membranes, VPU and β-tubulin proteins were revealed by immunoblotting with polyclonal antibodies followed by chemilumenescence detection. C. 293T cells were transfected with an HIV-1 LTR-GFP construct that does not encode Tat along with TASK-3, Vpu or empty vector. After 48 hours, the cells were harvested and subjected to flow cytometry analysis for GFP expression. Data shown are mean fluorescence intensity of transfected cells. Non-transfected cells were used to set the negative gate. (* P < .001; Student T test was used to determine statistical significance).