FIG. 4.

Characterization of the infectivity and nuclear import abilities of mutant virus lacking all potential karyophilic elements. (A) Quantities of wild type and ΔNLS (MA NLS⁻, IN 1-50, Vpr⁻, and ΔcPPT-CTS) mutant viral particles released in the supernatant were determined by p24CA ELISA after purification. (B) Normalized quantities of particles were analyzed by Western blotting using the indicated antibodies. (C) Virions were used to infect cycling and aphidicolin-treated HeLa cells, PHA-IL-2-stimulated PBLs, macrophages, and DCs. The percentages of infected GFP-positive cells were assessed 3 to 5 days following infection by flow cytometry and are presented here normalized to the WT level. (D and E) PCR analysis was carried out as described for Fig. 2, and the relative quantities of FL products of reverse transcription (black bars) and the 1-LTR-circle/FL-DNA (white bars) and 2-LTR-circle/FL-DNA (gray bars) ratios were determined by PCR. The results presented for viral DNA circle species are normalized to the amount of FL DNA synthesized for each mutant, as indicated. Each graph presents values obtained in 2 to 4 experiments. Values that are significantly lower than those observed for the wild type are indicated by an asterisk (Student test; P < 0.05).