Figure 3

O-FISH detection of HIV-1 RNA genome. For HIV-1 nucleic acid detection, MT-2 cells were fixed 6 hrs post mock-infection (a), or infection with HIV-1 (b), onto glass slides and incubated with O-FISH pol probe targeted towards HIV-1 positive sense genomic RNA. Bound probe was detected using the O-FISH protocol. O-FISH signals are shown in red and nuclei in blue. Images were derived from a volume compression of a z-stack of 16 images taken at a 0.4 μm step size. Scale bars, 10 μm (a, b). MT-2 cells were infected with a mixture of equal amounts of HIV^GFP-Vpr and HIV^mCh-Vpr (negative co-localization control), fixed after 6 hrs of synchronized infection and imaged (c, d, e). HIV^GFP-Vpr is shown in green (c, e) and HIV^mCh-Vpr is shown in red (d, e) and a merged HIV^GFP-Vpr and HIV^mCh-Vpr image is shown (e). The provided image was derived from a volume compression of a z-stack of 25 images taken at a 0.5 μm step size. Scale bars, 5 μm (c, d and e). Nuclei were labelled with Hoechst 33258 (blue). All micrographs are representative of at least 5 images per condition.