FIG. 5.
Effect of bafilomycin A1 on HIV replication and entry into macrophages. (A) Single-cycle replication assay. Macrophages were preincubated with or without bafilomycin A1 (0.5 μM) and exposed to either HIV(VSV) (left) or HIVNLAD8 (right ). The virus inoculum corresponded to 50 ng of p24 for 4 h at 37°C. Macrophages were washed to eliminate drug and virus; 20 h after viral exposure, AZT was added to the culture medium to prevent secondary replication cycles. Virus replication was assessed by measuring p24 production in the cell supernatants. Data are means ± SD of triplicate measurements and are representative of at least three independent experiments. (B) Intracellular p24 levels in cytosolic and vesicular fractions. Macrophages were preincubated with or without bafilomycin A1 (0.5 μM) and exposed to HIV(VSV) (left ) or HIVNLAD8 (right ). Viral input corresponded to 450 ng of p24 for 2 h at 37°C. p24 contents in subcellular fractions were then measured. Values represent relative p24 levels in treated cells, with 100% corresponding to untreated cells. Data are means ± SD of at least three independent experiments.