Figure 2

Functional interplay between HIV-1 Vpr and HIF-1α. A and B. 786-O cells were transfected with LTR-Luc full length along with an increasing concentration of HIF-1α (A) and/or Vpr (B) expression plasmids. The amount of DNA in each transfection mixture was normalized with pcDNA₆HisA. Luciferase activity was determined 48 hours after transfection. Results are displayed as histograms. C. Approximately 100, 000 cpm of synthetic [γ³²P]-labeled double-stranded DNA oligonucleotide probe corresponding to the HIV-1 LTR GC-rich site was incubated with 10 μg of nuclear extracts prepared from microglial cells transfected with Vpr expression plasmid. Labeled probe was also incubated with nuclear extracts prepared from pcDNA-transfected microglial cells in the presence of anti-Vpr antibody (lane 4) and normal mouse serum (NMS) (lane 2). D. To further examine the interplay between Vpr and HIF-1α, 786-O cells were transfected with LTR-Luc along with HIF-1α, Vpr and/or Sp1 using different combinations as shown. Luciferase activity was determined 48 hours after transfection. Results are displayed as histogram. Each transfection was repeated 3 times using different DNA preps.