Skip to main content
. 2019 Sep 13;13:399. doi: 10.3389/fncel.2019.00399

FIGURE 5.

FIGURE 5

The miR-204-5p directly targets the 3′UTR of DYRK1A. (A) WT DYRK1A 3′UTR contains three predicted binding regions of miR-204-5p. Mut1, Mut2, Mut3, or Mut4 DYRK1A 3′UTR was prepared for analyzing the interaction between miR-204-5p and DYRK1A 3′UTR. (B) Dual-luciferase reporter assay was used to evaluate the interaction between miR-204-5p and DYRK1A 3′UTR. SH-SY5Y cells were cotransfected with reporter vector containing WT or mutant DYRK1A 3′UTR along with scramble miR control (SC) or miR-204-5p mimic. The level of luciferase activity was significantly increased in SH-SY5Y cells cotransfected with WT DYRK1A 3′UTR and miR-204-5p mimic. Compared with SH-SY5Y dopaminergic cells cotransfected with WT DYRK1A 3′UTR and miR-204-5p mimic, the luciferase activity of SH-SY5Y cells cotransfected with Mut1, Mut2, or Mut3 DYRK1A 3′UTR and miR-204-5p mimic was decreased. The miR-204-5p mimic failed to affect the luciferase activity of SH-SY5Y cells cotransfected with Mut4 DYRK1A 3′UTR. For dual-luciferase reporter assays, each experiment was performed in triplicate. Each bar represents the mean ± SEM value of four independent experiments. ∗p < 0.05, ∗∗p < 0.01 compared with scramble miR control (SC)-transfected SH-SY5Y neurons expressing WT or mutant DYRK1A 3′UTR. #p < 0.05, ##p < 0.01, ###p < 0.001 compared with SH-SY5Y cells cotransfected with WT DYRK1A 3′UTR and miR-204-5p mimic.