Fig. 5.

Antiviral activity of RNA exosome. (A and B) HeLa cells were transfected with expression vectors containing EXOSC1, EXOSC4, and EXOSC5, and 24 h later, the transfected cells were infected with VSV at an MOI of 1 or 0.5. One day after infection, the cells were fixed and stained with crystal violet (A). Viral titers of culture media after 76 h were measured by a plaque assay (B). (C) Expression of EXOSC4 and EXOSC5 in stable HeLa clones, which express shRNA for EXOSC4, EXOSC5, or GFP, was examined by RT-PCR (upper panel), RT-qPCR (middle panel), and Western blotting (lower panel). The amounts of EXOSC4 and EXOSC5 cDNA in each sample were normalized by dividing by the amount of GAPDH. (D and E) Cell growth rates of stable HeLa clones, which express shRNA for EXOSC4, EXOSC5, DDX60, or GFP, were determined (D). The cells were infected with VSV at an MOI of 0.1 for 48 h, and viral titers of culture media were determined by a plaque assay (E). (F) FLAG-tagged EXOSC4- and HA-tagged DDX60-expressing vectors were transfected into HEK293FT cells. After VSV or mock infection, the immunoprecipitation was performed with anti-HA antibody.