Figure 2

TASK proteins suppress HIV-1 transcription. A. 293T cells were transfected with pNL4-3Luc and TASK-1 expression vector or empty vector (EV). After 48 hours cells were lysed and a luciferase assay was performed to determine the effect of TASK-1 on HIV-1 transcription. Data are normalized to the empty vector control (EV). B. Transfected 293T cells were harvested and lysed with a detergent buffer containing protease inhibitors. Equal protein amounts of cell lysate were separated by 4-12% SDS PAGE and transferred to nitrocellulose membranes. TASK and β-tubulin proteins were revealed by immunoblotting with polyclonal antibodies followed by chemilumenescence detection. C. 293T cells were transfected with pNL4-3Luc and 0.5ug, 1ug, or 2ug TASK-1 or empty vector. After 48 hours cells were lysed and a luciferase assay was performed. Data are normalized to the empty vector control (EV). D. Transfected 293T cells from (C.) were harvested, lysed and equal amounts of protein from cell lysates were subjected to 4-12% SDS PAGE. After transfer to nitrocellulose membranes, TASK and β-tubulin proteins were revealed by immunoblotting with polyclonal antibodies followed by chemilumenescence detection. (* P < .0001; Student T test was used to determine statistical significance).