FIG 5.

Correlation of small fluorescent HIV Gag-containing puncta with electron-dense virus-like particles within enclosed membrane compartments. (A) FIB-SEM photo showing one donor cell, labeled D, and three target cells, labeled T. Reconstructed nuclei are shown in transparent red, and small GFP puncta are shown in segmented yellow. The target cell in the middle with segmented yellow signals is the same cell shown in Fig. 3. (B) 3D side view of an FIB-SEM section and segmented small GFP puncta. A red arrow indicates the plane of intersection of this FIB-SEM section with the 3D representation of the segmented puncta. (C) Front view of the section illustrated in panel B with location of the registered small GFP puncta. A red square marks the enlarged area magnified in the inset. Bar, 1 μm. (D) A section from the AMD-3100-untreated sample in shown in Fig. 4A. A red square marks the area magnified in the inset. Bar, 1 μm. (E) A volume around the intersection area as shown in panel C is segmented to highlight 3D membrane structures that contain virus-like particles. (F) A volume around the intersection area as shown in panel D is segmented to highlight 3D membrane structures that contain virus-like particles. For panels E and F, plasma membrane is shown in white, nuclear membrane is in red, intracellular vesicles are shown in magenta), and virus particles are shown in yellow (panel E) and green (panel F).