Analysis of MAVS dimerization and inhibition by HCV NS3/4A in HEK293 cells.

<p>(<b>A</b>) Evaluation of the HAVI-Fluc-MAVS association with Rluc-MAVS using the DLR-PD assay. HEK293 cells were co-transfected with the indicated vectors (100 ng). The percentage of Rluc on the beads compared with the cell lysate was calculated by dividing the Rluc activity of Rluc-MAVS on the beads with the Rluc activity in the same amount of lysate used in the pull-down assays. The Rluc/Fluc ratio on the beads was calculated by dividing the Rluc activity by the Fluc activity measured on the beads. (<b>B</b>) HAVI-MAVS was cleaved by HCV NS3/4A and detected by Western blot when coexpressed in HEK293 cells. (<b>C</b>) Analysis of the interactions between HAVI-Fluc-MAVS and Rluc-MAVS inhibited by HCV NS3/4A using the DLR-PD assay. HEK293 cells were co-transfected with HAVI-Fluc-MAVS (100 ng), Rluc-MAVS (100 ng) and DNA (100 ng) containing the NS3/4A vector (0, 10, 50 or 100 ng) and control plasmid pCI neo. The Rluc/Fluc ratio on the beads was calculated by dividing the Rluc activity by the Fluc activity measured on the beads.