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. 2018 Mar 8;19(3):773. doi: 10.3390/ijms19030773

Figure 3.

Figure 3

miR-25-3p directly targets the 3′UTR of Akt1. (A) The sequences of miR-25-3p target elements in the Akt1 3′UTR were relatively conserved in many mammals. These sequences were obtained from TargetScan. (B) Site-directed mutagenesis of the miR-25-3p target site in the Akt1 3′UTR; mutated bases shown in red. (C) Dual luciferase reporter assay. The Akt1 3′UTR/mutant plasmid was cotransfected with miR-25-3p mimics/NC, respectively, into growing C2C12 cells; dual luciferase activities were measured from cell lysates (24 h after transfection). miR-25-3p mimics/NC or inhibitors/NC were respectively transfected into growing C2C12 cells. After 48 h, Akt1 expression was detected by qRT-PCR (D) and Western blotting (E). NC = negative control (miR-239b-5p of caenorhabditis elegans). β-actin served as the loading control. Data were presented as means ± SD (n ≥ 3). ** p < 0.01; NS, not significant.