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. 2006 Dec 5;4(12):e435. doi: 10.1371/journal.pbio.0040435

Figure 10. Actin Influences HIV-1 Gag Localization but Not Particle Release in Infected Macrophages.

Figure 10

(A) Immunofluorescent detection of HIV-1 CA in macrophages, 38 h after infection by pseudotyped NL4–3 (Env−). Deconvolved optical sections acquired at the center of the vertical dimension of the cell, and at the cell–coverslip interface are shown for examples of infected cells cultured in the presence of DMSO or cytochalasin D. Cells were stained with anti–HIV-1 CA antibodies (red) and Hoechst 33258 (blue). Inset shows expanded view at the PM–coverslip interface and individual Gag puncta. Scale bar indicates 10 μm.

(B) Quantitative analysis of Gag localization in NL4–3 (Env−) infected macrophages. The proportion of cells exhibiting each Gag distribution among 44 to 49 cells counted is plotted.

(C) Distribution of Gag in NL4–3 (MA/YFP) infected macrophages, as in (A) except that no immunostaining was done. Inset shows expanded view at the PM–coverslip interface and individual Gag (MA/YFP) puncta. Scale bar indicates 10 μm.

(D) Western blot analysis of HIV-1 release from NL4–3 (Env−) infected cells in the presence or absence of cytochalasin D, as indicated. Numerical values below the blots indicate p24CA signal intensities in cells and VLPs, derived by densitometry.