FIG. 6.
Comparative analysis of the synthesis of viral DNA in microglia infected with a panel of HIV-1 isolates. Adherent layers of microglia were infected with the panel of HIV-1 isolates, and at 24 and 72 h postinfection, supernatant fluids were collected for analysis of RT activity and the cells were collected for extraction of cellular DNA for PCR analysis. Early, intermediate, and late products of RT were amplified as described in Materials and Methods. Episomal forms of HIV-1 DNAs were detected with primers to gag and nef and LTR R/U5 regions. The amplified products were analyzed by electrophoresis on agarose gels and then hybridized to radiolabeled oligonucleotides to detect specific regions of HIV-1 proviral DNA. The accumulated viral DNA products were then quantified on a PhosphorImager. HIV-1DJV (DJV)-, HIV-1BAL (BAL)-, and HIV-189.6 (89.6)-infected microglia (A) and HIV-1JR-FL (JR-FL)- and HIV-1SF162 (SF-162)-infected microglia and infected microglia (B) are shown.