Fig. 3.

VP56 locates in the cytoplasm and interacts with the RLRs. (A) HEK 293T cells seeded into 10-cm² dishes were transfected with empty vector or Flag-MAVS/TBK1/MITA/IRF3/IRF7 and Myc-VP56 (5 μg each). After 24 h, cell lysates were IP with anti-Flag affinity gel. The immunoprecipitates and WCLs were then analyzed by IB with anti-Flag and anti-Myc Abs, respectively. (B) EPC cells seeded on microscopy cover glass in 6-well plates were transfected with 2 μg of VP56-EGFP and 2 μg of empty vector or DsRed-MAVS/TBK1/MITA/IRF3/IRF7. After 24 h, the cells were fixed and subjected to confocal microscopy analysis. Green signals represent overexpressed VP56 protein, red signals represent overexpressed MAVS, TBK1, MITA, IRF3, or IRF7, and blue staining indicates the nucleus region (a × 63 oil immersion objective). Scale bar, 10 μm. All experiments were repeated at least three times with similar results. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)