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. 2015 Dec 3;5:17554. doi: 10.1038/srep17554

Figure 1. ORF4b predominantly localizes to the nucleolus and inhibits the induction of the IFN-β in a dose-dependent manner.

Figure 1

(A) Nuclear localization of ORF4b protein. HeLa cells were transfected with an expression plasmid for HA-tagged ORF4b protein, and then stained for ORF4b with anti-HA antibody (green) at 24 h post-transfection. Nuclei were stained with DAPI (blue). Cells were analyzed by confocal microscopy using a 100× objective, and representative images are shown. Scale bar, 20 μm. (B) ORF4b could not induce apoptosis. HeLa cells were transfected with an expression plasmid for HA-tagged ORF4b protein, and cell lysates were analyzed by Western blotting with anti-Caspase-3 and anti-PARP antibodies. FL indicates full-length and CF indicates cleaved fragment. (C) expression of ORF4b protein in cultured cells. The 293T cells were transfected with increasing amounts of an expression plasmid for HA-tagged ORF4b protein, and cell lysates were analyzed by Western blotting. (D) ORF4b inhibits the induction of IFN-β in a dose-dependent manner. 293T cells were co-transfected with pGL3-IFNβ-luc, the internal control pRL-SV40, and increasing amounts of plasmid expressing ORF4b. At 24 h post-transfection, cells were infected with Sendai virus, then cells were harvested at 24 h post-infection and analyzed for Firefly and Renilla luciferase. Data are representative of three independent experiments with triplicate samples.