A3A is resistant to Vif induced degradation. (A) HeLa cells were transfected with vectors expressing vif-deficient pNL4-3 (3 μg each) along with pcDNA-A3A (1.5 μg each) and 1.5 μg of either pNL-A1vif(-) (lane 1), pNL-A1 (lane 2), or pcDNA-hVif (lane 3). Cells were harvested 24 h after transfection and whole-cell lysates were analyzed by immunoblotting using an A3G-specific rabbit polyclonal antibody (ApoC17) followed by incubation with an HRP-conjugated anti-rabbit antibody (A3A). The same blot was subsequently re-blotted with a Vif-specific monoclonal antibody (Vif) followed by probing with an HIV-positive patient serum to identify capsid protein (CA). Proteins are identified on the right. (B) Virus-containing supernatants from panel A were normalized for equivalent amounts of reverse transcriptase activity and used to infect LuSIV indicator cells  for determination of viral infectivity as described in Materials and Methods. Luciferase activity induced by virus produced in the absence of Vif and A3G was defined as 100% (lane 1). The infectivity of the remaining viruses was calculated relative to the control virus. Error bars reflect standard deviations from triplicate independent infections.