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. 2010 Dec 8;5(12):e14258. doi: 10.1371/journal.pone.0014258

Figure 5. Properties of a 1b-type core protein in the IPS-1 pathway.

Figure 5

(A) A core protein derived from an HCV patient suppressed DDX3-mediated activation of IPS-1 signaling. The 1b-type core protein was cloned into the pCMV vector from a patient with hepatitis C. IPS-1 (100 ng), DDX3 (100 ng) and HCV core (100 ng) expression vectors were transfected into HEK293 cells with a reporter plasmid (p125luc), for analysis as in Figure 4. (B) The core protein reduced interaction between full-length DDX3 and IPS-1. The plasmids encoding core protein (400 ng), DDX3-HA (400 ng) and FLAG-IPS-1 (400 ng) were transfected into HEK293FT cells. After 24 hrs, cell lysates were prepared and immunoprecipitation was carried out using anti-HA (DDX3-HA). (C) The core protein blocked interaction between the C-terminal fragment of DDX3 and IPS-1. The C-terminal region of DDX3 (199–662 aa) called DDX3 2-3c, IPS-1, HCV (O) and JFH1 or 1b core expression plasmids were transfected into HEK293FT cells. After 24 hrs, cell lysates were prepared and immunoprecipitation was carried out with anti-HA (DDX3 2–3c). Immunoprecipitates were analyzed by SDS-PAGE and Western blotting with anti-HA or FLAG antibodies. The results are representative of two independent experiments.