Figure 5

Analysis of CA accumulation at the VS. Confocal microscopic analysis of CA and F-actin localisation phenotypes in conjugates of non-permissive H9 cells or permissive MT-4 cells infected with HIV-Wt, HIV-Env-Tr712 or HIV-ΔEnv. Conjugates between infected donor cells and freshly added dye-labelled target cells were generated as described in the Material and Methods section and selected for analysis initially by widefield microscopy. A. Predominant CA localisation patterns in conjugates of H9 (np) cells weakly infected with either HIV-Wt (distinct accumulation at the cell contact site), HIV-Env-Tr712 or HIV-ΔEnv (both diffuse cytoplasmic staining) as indicated. Dye-labelled cells, which were not visualised by our confocal microcope, are marked with × in the merge. B: Predominant CA localisation patterns in conjugates of weakly infected MT-4 (p) cells (CA accumulation at multiple sites at the cell periphery). Note that due to enhanced per cell CA expression levels in MT-4 cells, exposure times for taking the micrographs in B were approximately half as long as those employed in A in order to allow detection of individual CA clusters in both cases. C. The percentages of HIV-Wt conjugates exhibiting CA accumulation at the contact site (in the case of permissive MT-4 cells with or without accumulation elsewhere at the cell periphery) was set at 100% and the percentages of HIV-Env-Tr712 and HIV-ΔEnv conjugates exhibiting this phenotype calculated relative to this. The mean percentages from several experiments in weakly infected H9 cells (np, low) or MT-4 cells (p, low) are given.