Figure 1.

The GluA2 3' untranslated region (UTR) has a functional miR-124 target site.

(A) Schema showing the position and conservation of the fully complementary miR-124 target site at the 5' end of the mouse GluA2 3' UTR. The seed region of miR-124 is shown in bold. Vertical bars depict Watson-Crick base pairs.

(B–D) Wild type and mutant reporter constructs were transfected into HEK293T cells with miR-124 mimics or mutant miR-124 mimics with two point mutations in the seed region (underlined). Luciferase activities are reported relative to reporter-only controls. (B) Transfection of the wild type reporter (“WT”) with miR-124 resulted in robust knockdown of luciferase activity while transfection with mutant miR-124 did not. Significance determined by one-way analysis of variance (ANOVA) with Bonferroni correction. (C) Transfection of miR-124 with a reporter lacking the miR-124 target site (“TD”) did not reduce luciferase activity. ^ site of deletion. Significance determined by two-tailed t-test. (D) Transfection of a mutant reporter with two point mutations in the miR-124 target site (“PM”) did not show reduced translation by miR-124. Translational reduction was restored when the mutant reporter was transfected with the complementary mutant miR-124. Significance determined by one-way ANOVA with Bonferroni correction. ***p<0.001; error bars show standard error of the mean (S.E.M.); N = 4 independent experiments per group.