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. 2002 Nov;76(21):11054–11064. doi: 10.1128/JVI.76.21.11054-11064.2002

FIG. 6.

FIG. 6.

Us11C does not displace MBP-3 from PKR. Shown are the effects of increasing concentrations of purified Us11C on MBP-3 interaction with PKR. PKR was immunoprecipitated from pcDNA3-PKR-FLAG-transfected HT1080 cells by using anti-FLAG-agarose. After changing to a low-salt buffer, PKR was eluted off the beads with excess FLAG peptide. Purified Us11C and then purified PKR were added to amylose resin-immobilized MBP-3. The proteins were incubated for 1 h and washed extensively with low-salt buffer. The proteins interacting with MBP-3 were analyzed by Western blotting for PKR using FLAG antibody. Lanes 1 to 6, PKR; lanes 2 to 6, 1.0 μg of purified MBP-3; lane 3, 0.1 μg of purified Us11C; lane 4, 0.3 μg of purified Us11C; lane 5, 1.0 μg of purified Us11C; lane 6, 2.4 μg of purified Us11C. The bottom panel shows the stripped Western blot reprobed with anti-PACT domain 3 antibody.