Effects of HCV on Basal and Tat-Induced HIV LTR Activation
Figure 2
HCV Core-mediated suppression of HIV LTR activation.
A dose-response experiment was performed in Huh7.5 cells for basal (A), as well as Tat-mediated HIV LTR activity (B). Huh7.5 cells (∼2×104 cells per well) were seeded in a 96-well plate and co-transfected with 100 ng of HIV LTR-B luciferase construct or the delNFkB construct (hatched bars) and 20 ng, 100 ng, or 500 ng of an HCV Core expression vector with or without 100 ng of the Tat expression vector. The pCI control vector was used to equilibrate the total amount of DNA per well as well as a negative control. Luciferase assay was performed at 48 hours post-transfection and expressed as relative luciferase activity. Similar experiments were performed in 293T (C) and Jurkat cells (D). For Jurkats, ∼2×106 cells were seeded per well of 24-well plate and co-transfected with 500 ng each of LTR-B or Tat and 250 ng, 500 ng, or 1000 ng for HCV Core using the transfection reagent TransIT-Jurkat (MIRUSBIO). White bars denote basal (no Tat) LTR activity, and black bars denote Tat-mediated LTR activation. A dose-response experiment with HCV NS3/4A was performed in Huh7.5 cells for basal as well as Tat-induced HIV LTR activation (E). The effect of HCV Core was tested in the presence or absence of HCV NS3/4A on basal (F) and Tat-induced LTR activation (G).