Identification of Small Molecules with Type I Interferon Inducing Properties by High-Throughput Screening
Figure 5
IFN-β induction by C3 in the presence of viral antagonist, NS1 and NS3/4A.
A Induction of IFN by C3 in the absence (white bars) and presence (dark bars) of influenza A/PR/8/34 virus. Cells (either 293T-FF or MDCK expressing the firefly luciferase under the control of the IFN-β promoter) were infected with influenza A/PR/8/34 virus at an MOI of 10 and 2 hrs later treated with C3 at the indicated concentrations. Twenty hrs post-treatment the luciferase activity in the cells was measured as an indirect read-out of the IFN levels. The maximum luciferase value obtained from the mock infected cells for each cell line was set to 100%. B. and C. Induction of IFN in the presence of plasmid expressed NS1 from influenza A/PR/8/34 virus or NS3/4A from HCV. 293T-FF cells were transfected with either GFP (white bars), NS1 (dark bars) or NS3/4A (hatched bars), 24 hrs later the media was removed and fresh media containing C3 (C) or Sendai Virus (D) added. The luciferase levels were measured 24 hrs later and the relative luminescence units (RLU) of the GFP transfected cells set to 100%. Error bars represent standard deviation of three replicates. The significance of the differences was calculated using a t-test, ns indicates not significant, **pvalue<0.01 and ***pvalue<0.001.