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. 2007 Jun 13;81(17):8953–8966. doi: 10.1128/JVI.00649-07

FIG. 6.

FIG. 6.

NS5A interacts with the death domain of MyD88 through the ISDR and inhibits recruitment of IRAK to MyD88. (A) The structure of NS5A and the MyD88 binding region are indicated at the top. MyD88-His was coexpressed with C-terminal deletion mutants of Flag-NS5A in 293T cells, immunoprecipitated (IP) with anti-His antibody, and immunoblotted (IB) with anti-Flag antibody (left). MyD88-His was coexpressed with Flag-NS5A deletion mutants (Δ240-280 or Δ280-300) in 293T cells, immunoprecipitated with anti-Flag antibody, and then immunoblotted with anti-His antibody (right). (B) Flag-NS5A was coexpressed with N-terminal or C-terminal deletion mutants of MyD88-His (ΔN1, ΔN2, ΔN3, ΔC1, ΔC2, or ΔC3) in 293T cells, immunoprecipitated with anti-His antibody, and immunoblotted with anti-Flag antibody. The structures of MyD88 and the deletion mutants and the NS5A binding region are indicated on the left. (C) Flag-MyD88 (left) or Flag-NS5A (middle) was coexpressed with IRAK-1-Myc or IRAK-4-Myc in 293T cells, immunoprecipitated with anti-Myc antibody, and immunoblotted with anti-Flag antibody. Flag-MyD88 and IRAK-1-Myc were coexpressed with Flag-NS5A in 293T cells, immunoprecipitated with anti-Myc antibody, and immunoblotted with anti-Flag antibody. The effect of the increase in Flag-NS5A expression on the interaction of MyD88 with IRAK-1 was examined by transfection with 0.1, 0.5, or 2 μg of Flag-NS5A expression plasmid (right).