Fibroblasts or EBV-B cells were left untreated or were treated with IFN-α2b (Schering), IFN-β (PBL Assay Science), or IFN-γ (Imukin, Boehringer Ingelheim) for the specified times before lysis. Cells were lysed in NP-40 lysis buffer (280 mM NaCl, 50 mM Tris [pH 8], 0.2 mM EDTA, 2 mM EGTA, 10% glycerol, 0.5% NP-40) supplemented with 1 mM DTT, PhosSTOP (Roche), and cOmplete Protease Inhibitor Cocktail (Roche). The protein lysate was subjected to SDS-PAGE, and the bands obtained were transferred to a nitrocellulose membrane, which was probed with unconjugated primary antibodies and secondary antibodies adapted for LI-COR. An anti-GAPDH antibody (Santa Cruz Biotechnology) was used as a loading control. For endogenous IFNAR1, we used an antibody recognizing the N-terminus of the IFNAR1 protein at a dilution of 1:1000 (64G12 custom antibody), whereas, for the protein overproduced following transfection, we used a polyclonal anti-IFNAR1 antibody recognizing the C-terminus of IFNAR1 (ab45172, Abcam). Antibodies against STAT1 (610116, BD Biosciences), p-STAT1 (562070, BD Biosciences), STAT2 (07-140, EMB), p-STAT2 (4441, Cell Signaling Technology), STAT3 (catalog 610189, BD Biosciences), p-STAT3 (562072, BD Biosciences), TYK2 (C8, sc-5271, Santa Cruz Biotechnology), and GAPDH (sc-47724, Santa Cruz Biotechnology) were purchased from commercial suppliers. The membrane was incubated overnight at 4°C with the primary antibodies. SuperSignal West Pico Chemiluminescent substrate (Thermo Fisher Scientific) was used to visualize HRP activity, and this signal was detected with an Amersham Imager 600 (GE Life Sciences). See complete unedited blots in the supplemental material.

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