Verification of IFN inhibitors ability to block IFN induction or IFN signaling.

<p>(a) Small molecules reported to inhibit the IKK2 (TPCA-1) and TBK1 (BX795, MRT6884, MRT6707) components of the IFN induction pathway were tested using the A549/pr(IFN-β).GFP reporter cell-line which contains an eGFP gene under control of the IFN-β promoter. The IFN induction pathway and hence GFP expression was activated by infection with a DI rich stock of PIV5VΔC. Effect of inhibitors at various concentrations was measured by monitoring fluorescence at 18 hours post-infection. (b) Small molecules reported to inhibit the JAK1 (Cyt387, AZD1480, Ruxolitinib, Tofacitinib) component of the IFN signaling pathway were tested using the A549/pr(ISRE).GFP reporter cell-line which contains an eGFP gene under control of an ISRE promoter. The IFN signaling pathway and hence GFP expression was activated by supplementing the cell supernatant with purified IFN. Effect of inhibitors at various concentrations was measured by monitoring fluorescence at 48 hours post IFN stimulation.