Figure 6.

RDUR regulates the expression of several critical antiviral genes in vitro and in vivo. (A) The levels of IFN-β, IFITM3, Mx1 and ISG15 were examined by qRT-PCR in A549 cells expressing shRNAs specifically targeting RDUR or luciferase (control) after infection with or without WSN. Plotted are the average levels from three independent experiments. Data are shown as means ± SD. **P < 0.01. (B) A549 cell lines stably expressing specific shRNAs targeting RDUR and luciferase (control) were infected with or without WSN and harvested at 16 hpi, followed by Western blotting with the indicated antibodies. Shown are representative data from three independent experiments with similar results. (C) The mRNA levels of IFN-β, IFITM3, Mx1, ISG15 and OAS2 in RDUR over-expression and empty vector (EV) control A549 cells infected with or without WSN were determined by qRT-PCR. Plotted are the average levels from three independent experiments. Data are shown as means ± SD. *P < 0.05, **P < 0.01. (D, E) The protein levels of IFN-β were determined by ELISA in the lungs of mRDUR knockout or WT mice infected with WSN or seasonal IAV (3 mice each group). Plotted are the average levels from three independent experiments. Data are shown as means ± SD. *P < 0.05, **P < 0.01. (F) RDUR knockdown and control A549 cells were infected with or without WSN, and Western blotting was performed to detect indicated proteins. (G) RDUR overexpression and EV control A549 cells were infected with or without WSN and examined by Western blotting with the indicated antibodies. (H) A549 cell lines stably expressing specific shRNAs targeting RDUR and luciferase (control) were infected with or without WSN and examined by Western blotting with the indicated antibodies. (I, J) mRDUR knockout and control mice were infected with WSN or CA04 influenza virus. Western blotting was performed by using indicated antibodies. Shown are representative immunoblots from three independent experiments.