Figure 2.

miR-124 Negatively Regulates the Expression of Akt1 and Akt2 by Directly Binding to Their 3′ UTRs

(A) The predicted binding sites of miR-124 in the wild-type 3′ UTRs of Akt1 and Akt2. Mutations in the 3′ UTRs are highlighted in red. (B) miR-124 negatively regulates the mRNA expression of Akt1 and Akt2 in NSCLC cells. The mRNA expression level of Akt1 and Akt2 was measured using qRT-PCR in stably expressing miR-124 H1299 cells and stably expressing miR-124-antisense H292 cells. (C) miR-124 negatively regulates the protein expression of Akt1 and Akt2 in NSCLC cells. The protein expression level of Akt1 and Akt2 was measured using western blot in indicated NSCLC cells that stably express miR-124 (H1299) or miR-124-antisense (H292). (D) 3′ UTR luciferase reporter assay for Akt1 and Akt2. Stably expressing miR-124 H1299 cells were transfected with indicated 3′ UTR luciferase reporter construct. After 48 hr of transfection, luciferase intensity was assessed. The data are presented as the mean ± SD from three independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001.