FIGURE 6.

Vpr-dependent down-regulation of MUS81-EME1 is independent of SLX4. A, HEK293T cells were transfected with MUS81 and EME1 plasmids, with increasing amounts of SLX4 plasmid along with Vpr or empty vector plasmid as indicated. Cells were harvested, and proteins levels were analyzed by immunoblotting with appropriate antibodies. B, SLX4 siRNA or control siRNA was transfected 24 h prior co-transfection with the indicated combinations of Vpr, MUS81, EME1, or balanced empty plasmid. C, purified MUS81·EME1, GST, GST-His₆-Vpr, GST-DDB1·DCAF1, and GST-DDB1·DCAF1·Vpr complexes were separated by SDS-PAGE and stained with Coomassie Blue. D, GST, GST-His₆-Vpr, GST-DDB1·DCAF1, or GST-DDB1·DCAF1·Vpr complex was incubated with recombinant MUS81-EME1. Bound fractions were analyzed by Western blotting. All experiments were repeated three times with similar results.