Figure 3

Overexpression of DDX1 in HeLa cells rescues the Rev/RRE-dependent reporter gene expression suppressed by Nullbasic. A. Schematic maps of pGag-RRE and pGag-CTE plasmids. B. Left: HeLa cells were transfected with appropriate expression plasmids, including pGag-RRE (500 ng), pRSV-Rev (10 ng), Nullbasic (NB, 500 ng), HA-DDX1(1000 ng), HA-DDX3 (1000 ng), DDX17 (1000 ng) and empty plasmid (pcDNA3-HA, 1000 g). Right: HeLa cells were transfected pGag-CTE (500 ng) with pRSV-Rev (10 ng), Nullbasic (NB) (500 ng), HA-DDX1(1000 ng) or empty vector (pcDNA3-HA, 1000 ng). A pcDNA3 plasmid was used to normalize the total amount of transfected plasmids and a luciferase expression plasmid was included to monitor transfection efficiency. After 24 h transfection, the cellular lysates were collected for assay of HIV-1 capsid level and luciferase activity. The p24 production was normalized to the luciferase reporter activity and expressed as a percentage relative to cells transfected with pGag-RRE with pRSV-Rev (left) or pGag-CTE with pRSV-Rev (right). Columns represent the means and standard deviations of transfections performed in triplicate. The experiment was performed 3 times with similar results.