Figure 5

Interaction of Cav-1 and Nef. U87-CD4-CXCR4 cells were co-transfected with pCZ-Cav-1 and HA tagged Nef (pCI NL4-3 Nef-HA-WT) or NefG2A(pCI NL4-3 NefG2A-HA). Twenty-four hours post transfection the cells were cultured in the presence of cholesterol (30 μg/ml) for 48 hours followed by apoA-I (20 μg/ml) for 10 min. (A) The cells were harvested and subjected to immunoprecipitation using anti-HA antibody and immunoblots using anti-Cav-1 or anti-Nef antibody. (B) Alternatively, immunoprecipitaion was performed using anti-Cav-1 and immunoblotting using anti-Nef antibody.