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

FIG. 2.

FIG. 2.

Us11C binds to PKR residues 1 to 170. GST pulldown assays were used to measure the binding of GST-tagged Us11C to PKR and PKR deletion mutants. FLAG-tagged PKR and PKR mutants were transfected into HT1080 cells, and cell extracts were prepared. The cell extracts were mixed with either purified GST or GST-Us11C protein in binding buffer, and GST-containing protein was pulled down using glutathione-Sepharose 4B. The PKR-FLAG protein constructs interacting with the GST-containing protein were analyzed by Western blotting with FLAG antibody. In panels B to D, the amount of extracts used was twice that used in panel A. (A) Input wt PKR and mutant proteins used to measure Us11C binding. The major band in each lane shows the position of the protein. (B to D) PKR mutant proteins interacting with GST-Us11C (B), GST (C), and GST-Us11N (D). Lanes 1, full-length PKR 1-551 (K296R); lanes 2, PKR 1-170; lanes 3, PKR 171-551; lanes 4, PKR 360-551.