Fig. 4

The block is post-entry at reverse transcription and independent of p21 and RNR. a Monocytes from 3 healthy donors (dot, square, triangle) were treated with the indicated R848 concentrations (blue) and infected (VprBlam) or uninfected (−) with two different amounts of Vpr.Blam-containing HIV-1. The ability of the cells to support virus fusion was measured by flow cytometry. b, c Monocytes from 3 healthy donors (one bar per donor) were treated with 10 µM R848 (blue), 100 U/mL IFNα or untreated (–) and 24 h later infected with Vpx-containing HIV-1 reporter virus (HIV1 X+). Uninfected (mock) or Nevirapine treated cells and cells infected with HIV-1 reporter virus lacking Vpx (HIV1) served as controls. 40 h post-infection, DNA of infected cells was isolated. Early (b) and late (c) HIV-1 reverse transcripts were quantified by RT-qPCR. d Monocytes from two donors (one bar per donor) were treated (+, blue) or untreated (−, grey) for 24 h with 10 μM R848. RNA was isolated and p21, RRM2 and IL-1β mRNA transcripts were quantified by qRT-PCR and normalized to GAPDH