Nef Alleles from All Major HIV-1 Clades Activate Src-Family Kinases and Enhance HIV-1 Replication in an Inhibitor-Sensitive Manner
Figure 8
Replication of HIV-1 Nef chimeras in CEM-T4 cells.
A) CEM-T4 cells (1×104 per well of a 96-well plate) were infected with 62.5 pg p24 equivalents/ml of wild-type HIV-1 NL4-3, a Nef-defective mutant (ΔNef), or the indicated Nef chimeras in a final culture volume of 200 µl. HIV p24 levels were determined by ELISA 10 days later. Data are presented as percent of p24 release observed relative to the NL4-3 control ± S.D. B) CEM-T4 cells (5×105 per well of a 6-well plate) were infected with 1 ng p24 equivalent/ml of the same panel of viruses as in part A. Viral Nef and gag protein expression was verified by immunoblotting lysates of infected cells 6 days later. Lysates from uninfected cells are included as a negative control (CEM-T4; far left lane).