NS1 interacts with the structural proteins.

Abstract

<p>(<b>A</b>) <i>Capsid</i>, <i>prM and Envelope proteins co-immunoprecipitate with NS1</i>. Naïve VeroE6 (CTRL), VeroE6_NS1^WT (WT) or VeroE6_NS1^HA (HA) cells were infected with 1 MOI of DVR2A^ΔNS1 TCPs. Forty-height hours post-infection, cell lysates clarified by centrifugation were used for immunoprecipitation with HA-affinity agarose beads and eluates (IP) or whole cell lysates (Input) analyzed by western-blotting using antibodies specified on the right of each panel. Numbers on the left refer to molecular weight standards expressed in kDa; black arrowhead on the right indicates the ΔNS1 protein expressed by the DVR2A^ΔNS1 genome. A representative experiment of four independent repetitions is shown. (<b>B</b>) <i>Interaction between NS1 and prM/E in DENV-2 wild-type virus-infected cells</i>. VeroE6 cells were mock infected or infected with DENV-2 at an MOI of 1. Forty-eight hours later clarified cell lysates were used for immunoprecipitation using a NS1-specific rabbit polyclonal antiserum or the corresponding pre-immune (PIS) antiserum and protein A-Sepharose beads. After extensive washing, eluted protein complexes were analyzed by western-blotting using polyclonal anti-NS1 and anti-prM or mouse monoclonal anti-E or anti-C specific antibodies as specified on the right of each panel. DENV proteins are indicated with arrowheads, asterisks refer to the immunoglobulin heavy chain.</p