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. 2020 Jun 15;11(6):463. doi: 10.1038/s41419-020-2625-7

Fig. 7. 5′ppp-RNA activation of RIG-I stimulates the IRF3–IFNβ–IFNAR–STAT1–IFIT1 pathway to induce cell death of A549 cells.

Fig. 7

Cells were transfected with Lyovec control or 1 μg/ml of 5′ppp-RNA with Lyovec. b, d, f Lysates were collected at indicated times after 5′ppp-RNA transfection and assayed for the indicated proteins. Blots are representative of three independent experiments. a, c, e, g Cell viability was determined using Cell Titer assay and normalized to the Lyovec control after 72 h. Specifically, cells were a treated with human recombinant IFNβ for the stated times, b, c pre-and cotreated with 100 or 200 μM Amlexanox, a TBK1 antagonist 1 h prior to 5′ppp-RNA transfection, d, e pre-and co-treated with 0.1 μg/ml B18R an IFNAR antagonist 1 h prior to 5′ppp-RNA transfection, and f, g transfected with negative siCTRL and siIFIT1 16 h post-transfection of 5′ppp-RNA. Data is represented as mean ± SEM of n = 3 independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001 vs. controls, #P < 0.05; ##P < 0.01; ###P < 0.001 vs. A549 parental cells using two-way ANOVA and Bonferonni post-tests.