Fig. 3

Generation of noninfectious virus by A1752. a A schematic representation of a reinfection experiment to examine infectivity of virus produced. b Comparison of antiviral activity of A1752 with other anti-HIV inhibitors. MT-4 cells were infected with HIV-1 NL4-3/EGFP virus in the presence of the indicated inhibitors. Viral production was determined using HIV-1 p24 ELISA. AZT, Tenofovir, DIBA, and SAMT were also used for comparison. c Determination of infectivity (Reinfection assay) of the viruses obtained in b. Fresh MT-4 cells were infected with the same amount (2.5 ng in p24) of purified viral particles produced from the infection assay in (b) in the absence of any inhibitors. Then, the GFP expressing cells were quantified using FACS analysis 48 h after the infection. d The viral (-)ssDNA, an early RT product, produced for 6 h after infection with viruses of each case from (b) was determined using qPCR. Data are the mean ± SEM of three separate experiments. e Determination of HIV-1 gRNA in MT-4 cells infected with an equal amount (30 ng in p24) of the virus particles produced from (b). At 3 h after infection, total cellular RNA was analyzed using northern-blot for detection of gRNA in the infected MT-4 cells. Gag-specific probe was for HIV-1 gRNA and an actin probe was used as a loading control