Fig. 2

Transfection of monocyte-derived macrophages (MDMs) with HIV-1 R3A WT, PTAP^– or Δp6 mutants by electroporation. Fourteen-day-old MDMs were electroporated with the indicated provirus constructs and incubated for 24 h before analysis. a Released virions were collected and virus pellets and cells were lysed and analysed by western blotting as detailed in Fig. 1b, and signal intensities were quantified using ImageJ to determine the virus release efficiencies shown below the blot (data from three independent experiments ± standard deviation). b Cells were fixed and stained with antibodies to p24/55 (38:96 K and EF7, green) and CD44 (magenta). Representative single confocal sections from z-stack series are shown. Scale bars, 10 μm. c Distributions of virus staining on the immunolabelled MDMs (samples from three different blood donors). Coverslips were scanned systematically and virus-expressing cells scored into three categories of cells where virus was seen in the intracellular plasma membrane-connected compartments (IPMCs) only (blue), in IPMCs as well as the cell surface (grey), or only at the cell surface (white; these latter MDMs lacked CD44⁺ IPMCs)