Figure 7

Suppression of HIV-1 transcription by Vpu and TASK-1 is abrogated by overexpression of RelA. Jurkat cells were transfected with EV, TASK-1 or Vpu or EV + RelA, TASK-1 + RelA or Vpu + RelA and then infected with wildtype NL4-3CFP (A) or NL4-3-D116N-YFP (B). Twenty four hours after infection the cells were lysed and total RNA was isolated and used as template for real-time PCR to measure nef mRNA. Data shown are normalized to empty vector control (C). Transfected Jurkat cells were harvested, lysed and equal amounts of lysate protein were subjected to 4-12% SDS-PAGE. After transfer to nitrocellulose membranes, Vpu, RelA and β-tubulin proteins were revealed by immunoblotting with polyclonal antibodies and visualized by chemilumenescence detection. (* P < .0001; Student T test was used to determine statistical significance).