FIG 4.
Mapping of STAT1 interacting domains in BoHV-1 VP8. HEK293T cells were transfected with plasmids containing FLAG, FLAG-VP8, or different N-terminally and C-terminally truncated FLAG-tagged VP8 versions as indicated. At 48 h posttransfection, cell lysates were made and incubated with anti-STAT1 antibodies (ab), followed by incubation with protein G-Sepharose. Immune complexes were separated by SDS-PAGE and detected by Western blotting. Immunoprecipitation of N-terminally (A) and C-terminally (C) truncated FLAG-tagged VP8 with anti-STAT1 antibody. Input lysates of cells transfected with N-terminally (B) and C-terminally (D) truncated FLAG-tagged VP8. (E) Immunoprecipitation of VP8 259-371, 372-483, 631-686, and 687-741 with anti-STAT1 antibody. (F) Input lysates of cells transfected with VP8 259-371, 372-483, 631-686, or 687-741. Truncated and full-length VP8 and STAT1 were detected using antibodies specific for FLAG and STAT1, respectively. It should be noted that anti-STAT1 antibody detects both STAT1α and STAT1β. Molecular wei markers (× 10−3) are indicated in the left margins.