Anti-BNRF1 was kindly provided by Dr. Lieberman [59 (link)], and anti-BZLF1 antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Polyclonal antibodies against BMRF1, BALF4 (glycoprotein B), and LMP1 were described previously [56 (link)]. Anti-STAT1 (#9172), anti-phospho-STAT1 (Tyr701) (#9171), and anti-GAPDH (#5174) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-STAT2 (#693302) and anti-phospho-STAT2 (Tyr631) (#619851) antibodies were purchased from Biolegend. Anti-HA antibody (3F10) (#11867423001)and anti-Flag antibody (M2) (F1804) were purchased from Sigma-Aldrich.
Immunoblotting was performed as described previously [60 (link)]. Densitometry was performed using ImageJ.
For surface staining, cells were incubated with anti-glycoprotein B antibody before fixation. Then cells were stained with Alexa 647-anti-mouse IgG (A-21235; ThermoFisher Scientific, Waltham, USA) on ice for 30 min. Antibody-stained cells were fixed overnight with 4% paraformaldehyde at 4°C. Subsequently, cells were treated with 0.1% Triton-X100/PBS at room temperature for 10 min. Cells were then stained further with PE-anti-BZLF1 antibody (sc-53904 PE; Santa Cruz Biotechnology) on ice for 30 min. Cells were analyzed using a BD Fortessa X-20.
Immunoblotting was performed as described previously [60 (link)]. Densitometry was performed using ImageJ.
For surface staining, cells were incubated with anti-glycoprotein B antibody before fixation. Then cells were stained with Alexa 647-anti-mouse IgG (A-21235; ThermoFisher Scientific, Waltham, USA) on ice for 30 min. Antibody-stained cells were fixed overnight with 4% paraformaldehyde at 4°C. Subsequently, cells were treated with 0.1% Triton-X100/PBS at room temperature for 10 min. Cells were then stained further with PE-anti-BZLF1 antibody (sc-53904 PE; Santa Cruz Biotechnology) on ice for 30 min. Cells were analyzed using a BD Fortessa X-20.
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