FIG. 6.

A3F expressed in stable HeLa cell lines does not have antiviral activity. (A) Stable HeLa cell lines expressing A3G wt (34), A3F wt (clones 1 and 2), or A3F mt (clones 1 and 2) were each transfected with 5 μg of pNL4-3 or pNL4-3Vif(−) DNA. Transfected cells and virus-containing supernatants were harvested 24 h later and processed for immunoblot analysis. Blots were probed with a myc-specific antibody (parts I and IV), a Vif MAb (part II), and an HIV-positive patient serum (APS) for the detection of viral CA proteins (parts III and V). Proteins are identified on the right. (B) Virus samples from panel A were normalized for equivalent reverse transcriptase activity and used to infect TZM-bl cells. Tat-induced luciferase activity was quantified and used to calculate relative viral infectivity. The infectivity of WT virus was defined as 100% for each producer cell type (lanes 1, 3, 5, 7, 9, and 11), and the infectivity of Vif-deficient viruses was calculated relative to that of the corresponding WT viruses (lanes 2, 4, 6, 8, 10, and 12). Error bars reflect standard deviations calculated from three independent infections.