Anti ie1

Immunoprecipitation (IP) experiments were performed by incubating cells lysates (0.5% NP40 lysis mix) with the respective antibody followed by protein A or G-agarose beads. The recovered proteins were resolved on an SDS-polyacrylamide gel (12.5%). For analysis of total cell lysates, cells were lysed in 1%SDS (in PBS) with three rounds of heating (95 °C)/scratching tube against a metal test tube rack and quantified using a Pierce BCA protein assay kit (Thermo Scientific). Equal amount of proteins (20–30 μg) were separated on a SDS-polyacrylamide gel (12.5%). The analysis of IE1 proteins required additional (3×) protein lysates. The proteins from the gels were then transferred to a PVDF membrane (0.45μm, Thermo Scientific) and the membrane was incubated in 5% non-fat drymilk/0.01% Tween-20/0.02% sodium azide in PBS followed by a primary antibody (anti-CD46 (2 μg ml⁻¹)), anti-IE1 (2 μg ml⁻¹) or anti-GAPDH (0.1 μg ml⁻¹) Millipore) and anti-rabbit or anti-mouse conjugated to HRP (1:20,000 or 1:10,000, respectively). Uncropped and unprocessed scans are included in the Source Data file.

HFF cells were lysed with radioimmunoprecipitation assay (RIPA) buffer (Sigma) containing a protease and phosphatase inhibitor cocktail (Pierce Biotechnology, Waltham, MA). Equal amounts of protein (20 to 40 μg, depending on the target protein) were resolved in 8 to 10% (wt/vol) SDS-PAGE and transferred to polyvinylidene fluoride (PVDF) membranes (PALL, Cortland, NY). The membranes were blocked with TBST (150 mM NaCl, 10 mM Tris-HCl, 0.1% [vol/vol] Tween 20, pH 8.0) containing 5% (wt/vol) skim milk and analyzed with the following primary antibodies: anti-YAP (1:1000, Santa Cruz Biotechnology, Santa Cruz, CA, sc-101199), anti-STING (1:500, R&D systems, Minneapolis, MN, MAB7169), anti-IE1 (1:1000, Millipore, Billerica, MA, MAB8131), anti-pp52 (1:1000, Virusys, Sykesville, MD, ICP36), anti-pp28 (1:1000, Santa Cruz Biotechnology, sc-69749), and anti-β-actin (1,10,000, Santa Cruz Biotechnology, sc-47778). All blots were incubated overnight with primary antibody at 4°C with horseradish peroxidase-conjugated anti-mouse or anti-rabbit secondary antibodies for 2 h at room temperature. The enhanced chemiluminescence system (Atto, Tokyo, Japan) and X-Omat film (Kodak, Rochester, NY) were used to visualize the protein bands.