FIG 6.

Mapping of the ZFP36L1-targeting regions/sequences in the JEV genome. (A) Basic ARE units (AUUUA) in the JEV genome. (B and C) Schematic diagram of the reporter RNAs (top panel). 293T/17 cells overexpressing EGFP and HA-ZFP36L1 were cotransfected with 5′-capped renilla luciferase (Rluc) flanked by JEV NS5 RNA and control Firefly luciferase (Fluc) RNA (bottom panel) (B) or 5′-capped Fluc fused with JEV 3′-UTR RNA and control Rluc RNA bottom panel) (C) for 24 h. Relative luciferase activity was measured by a dual-luciferase reporter assay. (D) The reporter RNAs containing WT or mutant AREs are shown. A solid box indicates wild-type ARE (AUUUA) and open box mutant ARE (AGGGA) of JEV 3′ UTR. 293T/17 cells expressing EGFP, WT, and different zinc-finger-defective forms of ZFP36L1 were cotransfected with 5′-capped Fluc JEV 3′ UTR RNA (WT and mutant) and control Rluc RNA. Relative luciferase activity was measured by a dual-luciferase reporter assay. Data are from three repeated experiments and shown as mean ± SD. Statistical significance was analyzed by a two-tailed Student’s t test. **, P ≤ 0.01; ***, P ≤ 0.001.