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. 2022 Oct 31;96(22):e01513-22. doi: 10.1128/jvi.01513-22

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FIG 7 — The inhibition of eIF4A3 on TBK1-IRF3 signaling. (A) The effect of eIF4A3 on the interaction between IRF3 and TBK1. Coimmunoprecipitation and immunoblot analysis of HEK293T cells transfected with Flag-TBK1, HA-IRF3, and Myc-eIF4A3 or sieIF4A3 HEK293T cells. The band intensities were quantified, and relative precipitated HA-IRF3/Flag-TBK1 or IRF3/TBK1 ratios are shown below. (B) Effect of eIF4A3 on the phosphorylation of IRF3 induced by TBK1. HEK293T cells were transfected with Flag-TBK1, HA-IRF3, and Myc-eIF4A3 to detect p-IRF3 at Ser 386 by Western blot analysis. The band intensities were quantified, and relative IRF3 levels (p-IRF3/IRF3) are shown below. (C) Identification of a binding site for the IRF3-eIF4A3 interaction. Coimmunoprecipitation and immunoblot analysis of HEK293T cells transfected with HA-eIF4A3, Flag-EGFP, Flag-EGFP-ΔIRF3 and Flag-IRF3 or HA-TBK1, Flag-EGFP, Flag-EGFP-ΔIRF3, and Flag-IRF3. (D) Effect of eIF4A3 on the dimer formation of IRF3 induced by TBK1. HEK293T cells were transfected with Flag-TBK1, HA-IRF3, and Myc-eIF4A3 to detect IRF3 dimer by native PAGE analysis; * represents the indicated protein.