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

Figure 4. HCV core protein inhibits DDX3 promotion of IPS-1-mediated IFN-beta induction.

Figure 4

(A) Expression plasmids for IPS-1 (100 ng), DDX3 (200 ng) and/or HCVO core (50 or 100 ng) were transfected into HEK293 cells in 24-well plates with reporter plasmids, and reporter activity was examined. (B) Expression plasmids for IPS-1 (100 ng), DDX3 (100 ng), and/or HCVO or JFH1 core (10, 25, 50 or 100 ng) were transfected into HEK293 cells, and reporter gene expression was analyzed. (C) IPS-1-, IKKepsilon- or NAP1-expressing plasmids were transfected into HEK293 cells with HCV JFH1 core-expressing plasmids (25 or 100 ng), for reporter gene analysis. (D) Plasmids for expression of FLAG-tagged IPS-1 (400 ng), HA-tagged DDX3 partial fragment (400 ng) and HCVO or JFH1 core (400 ng) were transfected into HEK293FT cells. 24 hrs later cells were lyzed and the lysate was incubated with anti-HA Ab for immunoprecipitation. The DDX3 (224-662)-bound IPS-1 was blotted onto a sheet and probed with anti-Flag Ab. Whole cell lysate was also stained with anti-tag Abs. (E) IPS-1 (100 ng), DDX3 (100 ng), JFH1 core (50 ng) and/or p125 luc reporter (100 ng) plasmids were transfected with HEK293 cells, with HCV 3′UTR poly-U/UC (PU/UC) RNA (25 ng), synthesized in vitro. Cell lysates were prepared after 24 hrs, and luciferase activities measured. One representative of at least three independent experiments is shown except for panel D, which is a representative of two sets of the experiments.