Fig. 4.

Insensitivity of the UL16-gEtail interaction to NEM treatment. (A) Insect cells infected with a recombinant baculovirus expressing UL16-GFP were either untreated or treated with NEM prior to detergent lysis. The lysates were incubated with approximately equal amounts of the indicated GST constructs (determined by Ponceau S staining [data not shown]) attached to glutathione beads. The bound proteins were collected by centrifugation, separated by SDS-PAGE, and analyzed by immunoblotting for GFP. (B) Equal aliquots of purified His₆-UL16 protein (1.2 μg) were incubated with approximately equal amounts of the indicated GST constructs as estimated from a Ponceau S-stained gel (data not shown), and bound proteins were analyzed by immunoblotting with antibody specific for the His₆ tag. The positions of the unmodified and NEM-modified forms of UL16 are indicated (arrowhead and arrow, respectively). The shift in migration is less obvious in panel A because of the larger mass of the UL16-GFP chimera.