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. 2001 Nov;75(22):11166–11177. doi: 10.1128/JVI.75.22.11166-11177.2001

FIG. 4.

FIG. 4

FIG. 4

HIV-1 entry analyzed by immunofluorescence and by cellular fractionation. (A) Confocal microscopy analysis of intracellular p24 after macrophage exposure to HIV-1. Macrophages were exposed to the indicated strains for 30 min at 37°C, washed, fixed, and labeled with anti-Gag MAbs. (Top) Immunofluorescence analysis. (Bottom) Phase contrast of the same fields. Noninfected (NI) macrophages were similarly stained as a negative control. (B) p24 levels in subcellular extracts of macrophages exposed to HIV-1. Macrophages were exposed to the R5-tropic HIVNLAD8 strain (left) or to HIV(VSV) pseudotype (right). Noninfectious HIV-1 particles devoid of envelope protein (HIVΔenv) were used as a control. Viral input corresponded to 450 ng of p24 for 2 h at 37°C. After viral exposure, cells were treated by pronase to eliminate virus adsorbed at the cell, and p24 contents were measured in the cytosolic and vesicular (pellet) fractions. Total intracellular p24 levels (in picograms) are indicated over the bars. The percentages of cytosolic and vesicular p24 are shown inside the bars. Data are the means of triplicate measurements (the standard deviation [SD] was below 10%) and are representative of at least three experiments.

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