Figure 8. Late Endosomes Are Not Productive Sites of HIV-1 Particle Formation in Primary Human Macrophages.
(A) Distribution of retargeted Gag-GFP proteins in macrophages. Cells expressing C2-Gag-GFP, FYVE-Gag-GFP, or PX-Gag-GFP (green) were fixed at 24-h post-transfection. Scale bars indicate 10 μm.
(B) FYVE-Gag-CherryFP targets late endosomes. Macrophages expressing FYVE-Gag-CherryFP (red) and CD63-GFP or Lamp1-GFP (green) were fixed at 24-h post-transfection and stained with Hoechst 33258 (blue). Insets are expanded views of individual accumulations of FYVE-Gag-CherryFP. Scale bar indicates 10 μm.
(C) Western blot analysis of macrophages expressing Gag-GFP WT or mutants. Samples were collected at 24-h post-transfection. Cell and virion lysates were probed with anti–HIV-1 CA antibodies. Numerical values below the blots indicate VLP signal intensities, derived by densitometry.