Fig. 4

ZIKV NS4A interacts with both the CL and TM domains of MAVS and prevents RIG-I from binding MAVS. a A schematic diagram of the MAVS protein and functional domains: CARD-like domain (aa 10 to 77), proline-rich domain (aa 103 to 173) and transmembrane domain (aa 514 to 535). b Co-IP and western blotting analysis of 293T cells transfected with Myc-tagged NS4A along with vectors expressing the indicated Flag-tagged MAVS truncation forms or full-length MAVS. Empty vector was used as a negative control. c 293T cells were co-transfected with Flag-tagged RIG-I and Myc-tagged NS4A or an empty vector control for 24 h. Whole cell lysates were subjected to immunoprecipitation using an anti-MAVS antibody and analyzed by western blotting for RIG-I (c). 293T cells were transfected with Myc-tagged NS4A or an empty vector control for 24 h later, transfected with 20 μg/ml of poly(I:C). Whole cell lysates were subjected to immunoprecipitation using an anti-MAVS antibody and analyzed by western blotting for TRAF6 (d), or TBK1 (e). The expression of precipitated proteins was determined by western blotting analysis using the indicated antibodies (lower panel). The data shown are representative of three independent experiments with similar results