FIG 1.

BGLF2 associates with Tyk2. (A) 293T cells were transfected with plasmids expressing V5-tagged BGLF2 or V5-tagged VZV ORF13 as a control, and 24 h later the cells were washed with PBS and lysed with radioimmunoprecipitation assay (RIPA) buffer in the presence of protease and phosphatase inhibitors. After centrifugation, the supernatant was immunoprecipitated with V5-agarose beads and analyzed with mass spectrometry. Protein identification was based on UniProt, and spectral counts (total number of fragmentation spectra that map to peptides of the protein) for BGLF2, NEK9, GMIP, and Tyk2 from cells expressing BGLF2 or VZV ORF13 are shown. 293T/17 SF cells were transfected with GFP, BGLF2, or BGLF2ΔC for 48 h, and cell lysates were immunoprecipitated with mouse anti-V5 (B) or rabbit anti-Tyk2 antibody (C). The precipitated immune complexes were immunoblotted with antibody to Tyk2, STAT1, STAT2, STAT3, and V5 for BGLF2 and BGLF2ΔC. Raji cells were electroporated with plasmids expressing GFP or BGLF2-V5, cell lysates were immunoprecipitated with Tky2 antibody (D) or mouse anti-V5 or anti-flag (M2) antibody (E), and immunoblotting was performed with antibody to Tyk2 or V5 for BGLF2. 293T-BGLF2-V5 cells were induced to express BGLF2 with doxycycline; cell lysates were immunoprecipitated with antibody to V5 for BGLF2 (F), or Tyk2 or IgG (G); and immunoblotting was performed with antibody to Tyk2, V5 for BGLF2, STAT2, or STAT3.