Figure 6. Analyses of HIV-rtTA-Tat^fs Genomic RNA Accumulation Complemented with Tat and VP35.

(A) HEK293T cells were transfected with HIV-rtTA-Tat^wt, HIV-rtTA-Tat^fs, and HIV-rtTA-Tat^fs in combination with Tat^wt or increasing amounts of VP35. Two to three days after transfection, virus production was measured via CA-p24 ELISA, and total RNA was isolated.

(B) Total RNA (10 μg) was run on a denaturing agarose gel, and after blotting, probed with a Nef-specific radiolabeled probe. Relative genomic RNA accumulation was quantified by phosphorimaging and is indicated below. The RNA accumulation of the HIV-rtTA-Tat^wt was set at 100%.