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. 2016 Jun 22;6:28481. doi: 10.1038/srep28481

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Copyright © 2016, Macmillan Publishers Limited

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(A,B) 293T cells were transfected with NS1 (NS1-HA or NS1-FLAG) or NS1-70 (NS1-70-HA or NS1-70-FLAG) expression plasmids or empty vectors for 30 h. The cells were then treated with TNF-α (10 ng/ml) for 30 min. Cells were lysed and subjected to immunoprecipitation (IP) using mouse anti-FLAG tag or rabbit anti-p65. Mouse (or rabbit) IgG was used as negative control. IP products and 5% input samples were resolved by immunoblotting (IB). Rabbit anti–p65 was used for detection of p65. For detection of FLAG-tagged and HA-tagged proteins, the indicated mouse Abs were used. One of three experiments is shown.