Rabbit anti viperin

Cells were trypsinized and pelleted at 800 × g for 5 min. Nuclear lysates were extracted using NE-PER reagents (Thermo Scientific). Equal amounts of cellular material were loaded into 4–20% acrylamide gels (Bio-Rad) and imaged using a ChemiDoc Imaging System (Bio-Rad). Protein was transferred to a nitrocellulose membrane for 60 min. PBS with 5% (w/v) non-fat dried milk and 0.1% Tween-20 were used to block for at least 1 h at 4°C. Primary antibodies were then incubated with the membrane overnight at 4°C. Antibodies used were rabbit anti-ETV7 (Sigma, HPA029033), rabbit anti-IFIT1 (Cell Signaling, D2X9Z), rabbit anti-IFIT2 (Proteintech, 12604–1-AP), mouse anti-IFITM1 (Proteintech, 60074–1-IG), mouse anti-FLAG (Sigma, F3165), rabbit anti-Lamin B1 (Cell Signaling, D4Q4Z), rabbit anti-Viperin (Cell Signaling, D5T2X), rabbit anti-GAPDH (Cell Signaling, 14C10), and mouse anti-αtubulin (Sigma, T5168). Membranes were washed five times in PBS with 0.1% Tween-20 and then an anti-rabbit-HRP (Thermo, A16104) or anti-mouse-HRP (Thermo, A16072) secondary antibody in 5% milk was added for 1 h. The membrane was then washed five times and Clarity or Clarity Max ECL substrate (Bio-Rad) was added before being exposed to film and developed.