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

FIG 9.

FIG 9

The inhibition of eIF4A3 on IFN-β does not rely on ATP binding, RNA-dependent ATPase, or RNA helicase activities. (A) Schematic diagram of the eIF4A3 truncated segments. (B) Effect of eIF4A3, eIF4A3-Nt, or eIF4A3-Ct on the IFN-β luciferase activity. HEK293T cells were transfected with the HA-eIF4A3, HA-eIF4A3-Nt, HA-eIF4A3-Ct, or vector (HA) and the IFN-β luciferase reporter, plus the pRL-TK. Then, 24 h after transfection, the cells were stimulated by SeV for 10 h and measured by the luciferase assays (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001; two-tailed Student’s t test). (C) Effect of eIF4A3, eIF4A3(DEAD-DQAD), or eIF4A3(Δ motif I) on the IFN-β luciferase activity. HEK293T cells were transfected with the HA-eIF4A3, HA-eIF4A3(DEAD-DQAD), HA-eIF4A3(Δ motif I), or vector (HA) and the IFN-β luciferase reporter, plus the pRL-TK. Then, 24 h after transfection, the cells were stimulated by SeV for 10 h and measured by the luciferase assays (*, P < 0.05; **, P < 0.01; ***, P < 0.001; ****; P < 0.0001; two-tailed Student’s t test). (D) Interactions between Flag-IRF3 and HA-eIF4A3, HA-eIF4A3(DEAD-DQAD), or HA-eIF4A3(Δ motif I). HEK293T cells were transfected with Flag-IRF3 and HA-eIF4A3, HA-eIF4A3(DEAD-DQAD), or HA-eIF4A3(Δ motif I), followed by lysing at 24 h posttransfection. A co-IP assay was carried out using anti-HA immunomagnetic beads, followed by a Western blot assay.