FIG 5.

BGLF2 induces BZLF1 expression in lymphoblastoid cell lines. Akata-EBV (A) or Raji-EBV (B) cells were transduced with a lentivirus expressing GFP, with lenti6-BGLF2 (expressing BGLF2), or with a lentivirus expressing shBGLF2. After 4 days, cell lysates were prepared and immunoblotted for detection of the EBV immediate early protein BZLF1, BGLF2, or actin. (C) Akata-EBV cells were transduced with a lentivirus expressing shRNA against BGLF2 (shBGLF2) under the control of a doxycycline (DOX)-inducible promoter and treated with or without rabbit anti-human IgG (10 μg/ml). After 2 days, cell lysates were collected and immunoblotted for BZLF1, the EBV early antigen BMRF1, or actin. (D) Raji-EBV cells were transfected with plasmids expressing two different BGLF2 shRNAs (targeting BGLF2 nucleotides 257 to 280 or 598 to 620) or a control siRNA pool. Two days later, cell lysates were treated with sodium butyrate (NaB) and 12-O-tetradecanoylphorbol 13-acetate (TPA) to induce EBV reactivation, and after 2 more days, cell lysates were prepared and immunoblotted for detection of BZLF1, p-p38, p38, or actin. (E) Raji-EBV cells were transfected with a plasmid expressing GFP, BGLF2, or BGLF2LL-AA, and after 3 days, lysates were immunoblotted for BZLF1, p-p38, p38, BGLF2, and actin.