FIG 7.

The CUL2 dominant negative mutant CUL2ΔNedd inhibits the function of BIV Vif. (A) The CUL2ΔNedd mutant blocked the BIV Vif-induced degradation of btA3Z2Z3. HEK293T cells in T25 flasks were cotransfected with 2 μg btA3Z2Z3 and 2 μg NL4-3ΔVif, along with 2 μg BIV Vif or a control vector (VR1012) and with or without 2 μg of the CUL2ΔNedd mutant. Cell lysates from transfected cells were prepared and separated by SDS-PAGE, transferred to nitrocellulose membranes, and probed with the indicated antibody. Ribosomal P protein was used as the sample loading control. (B) The CUL2ΔNedd mutant could not inhibit btA3Z2Z3 virion packaging. Viruses from supernatants were analyzed by immunoblotting with antibodies against HA and p24. (C) The CUL2ΔNedd mutant could inhibit the infectivity of NL4-3ΔVif virus in the presence of BIV Vif and btA3Z2Z3. Viruses from the supernatants were analyzed as described in the legend to Fig. 1A. Errors bars represent the SDs for triplicate wells within one experiment.