FIG. 4.

TTV ORF2 protein interacts with IKKα and IKKβ. HeLa cells were transfected with either the empty vector pcDNA3.1(−) or pcDNA3.1(−)-TTV2. At 48 h posttransfection and after being treated with TNF-α (10 ng/ml) for 4 h, cell lysate was prepared for immunoprecipitation with monoclonal anti-IKKα, anti-IKKβ, or anti-IKKγ antibodies, with immunoglobulin G (IgG) taken as a control. Immune complex captured by protein A-Sepharose was separated by SDS-PAGE and analyzed by Western blotting with anti-TTV ORF2 antibody. One-tenth of the total cell lysates used for immunoprecipitation was loaded as a positive control for IKKα, IKKβ, and IKKγ. Results are representative of three different experiments. IP, immunoprecipitation; IB, immunoblotting.