Figure 4

klotho is a direct target of miR-199a-5p. (A) Profile of pYr-MirTarget-Klotho-3′UTR luciferase report plasmid; (B) The positions of the two miR-199a-5p target sites in klotho 3′UTR, showing sequence alignment of miR-199a-5p and the klotho 3′UTR; (C) Luciferase activity was dramatically decreased in the presence of miR-199a-5p mimic when compared with negative control (*P < 0.05). Renilla luciferase activity of each sample was normalized by Firefly luciferase activity. Data were shown as mean ± standard deviation from three independent experiments. (D) HEK293 cells were co-transfected with luciferase reporter plasmid containing either wild type or mutant klotho 3′UTR (indicated as klotho-Wt or klotho-Mut), and the luciferase activity of wild type klotho became much lower than that of mutant type klotho and negative control when miR-199a-5p was introduced (*P < 0.05).