Figure 3.
Premature reverse transcription can be blocked. HIV-1 was expressed from 293T cells transfected either in the absence (black bars) or presence (red bars) of RTIs. Virus was harvested and treated with DNase I and subtilisin, as described in the Methods section (Figure 1, right). Quantities of intravirion DNA were then measured by qPCR using the reverse transcription intermediate targets [31] indicated at the bottom of the figure (tRNA, red line; minus-strand DNA, black line; plus-strand DNA, blue line; target sequences indicated by the black dumbbells). The quantities expressed are the ratio of vDNA to gRNA. Panel A shows wild-type virus, panel B shows NCH23C virus, and panel C shows NCH44C virus. Values plotted are the means and the errors bars are the standard deviations from two separate experiments.