Fig. 4.

Analysis of engagement of Vpr M/D-lo and Vpr P to the DDB1-CUL4A (VprBP/DCAF-1) E3 ubiquitin ligase. HEK293 T cells were transfected with plasmids expressing Vpr (D-sh, D-lo, P, or HxB) or no Vpr (WPI). Cells were transcomplemented with plasmids expressing myc-VprBP (lanes 2, 4, 6, 8, and 10) or a myc tag alone (lanes 1, 3, 5, 7, and 9). Immunoprecipitation (IP) of total cell lysates was performed using anti-myc antibody. Resulting immunocomplexes (and total lysates, as a control) were analyzed for myc-VprBP, DDB1, and Vpr by Western blotting using the respective antibodies. The asterisk indicates an unidentified species having a higher molecular size than the expected size (∼14 kDa) for Vpr P. Shown are data representative of 3 experiments.