FIG 2.
BoHV-1 VP8 interacts with STAT1. (A to C) HEK293T cells were transfected with pFLAG-EYFP or pFLAG-VP8. (A and B) At 48 h posttransfection, cell lysates were generated and incubated with anti-FLAG resin (A) or anti-STAT1 antibody (B) followed by protein G-Sepharose. (C) Input lysates of mock-, pFLAG-EYFP-, and pFLAG-VP8-transfected cells that were used in the immunoprecipitation assays illustrated in panels A and B. (D to F) MDBK cells were mock infected or infected with BoHV-1 or BHV1-ΔUL47. (D and E) At 24 h postinfection, cell lysates were made and incubated with anti-STAT1 antibody (D) and anti-VP8 antibody (E), followed by protein G-Sepharose. (F) Expression of VP8 in BoHV-1-infected cells and STAT1 in mock-, BoHV-1-, or BoHV1-ΔUL47-infected cells. (G and H) HEK293T cells were transfected with pFLAG-EYFP or pFLAG-VP8. (G) At 48 h posttransfection, cell lysates were collected and incubated with anti-FLAG resin. (H) Input lysates of mock-, pFLAG-EYFP-, and pFLAG-VP8-transfected cells that were used in the immunoprecipitation assay illustrated in panel G. VP8, EYFP, STAT1, and STAT2 were detected by Western blotting with monoclonal anti-VP8 and anti-FLAG antibodies and rabbit anti-STAT1 and anti-STAT2 antibodies, respectively.