Figure 3
MiR-324-5p participates in the regulation of Mtfr1 expression. (a) MiR-324-5p levels were analyzed by qRT-PCR. *P<0.05 versus control. (b) Analysis of Mtfr1 3′UTR potential binding site for miR-324-5p. (c) Knockdown of miR-324-5p induces an elevated Mtfr1 levels. Cardiomyocytes were transfected with antagomir miR-324-5p (anta-324-5p) or antagomir-negative control (anta-NC). The expression of Mtfr1 was detected by immunoblot. (d) miR-324-5p attenuates A/R-induced Mtfr1 upregulation. Cardiomyocytes were transfected with miR-324-5p mimic (mimic-324-5p) and its negative control (mimic-NC), and then were treated with A/R. Mtfr1 levels were detected by immunoblot. (e) miR-324-5p binding site was mutated in the Mtfr1 3′UTR. (f) miR-324-5p inhibits the luciferase activity of Mtfr1 with wild-type 3′UTR. HEK293 cells were transfected with Mtfr1 with a wild-type 3′UTR or mutated 3′UTR, miR-324-5p mimic or its negative control, and then the cells were harvested, and luciferase activity was measured; *P<0.05. (g and h) miR-324-5p inhibits the expression of Mtfr1 with wild-type 3′UTR. Cardiomyocytes were infected with adenoviral Mtfr1 with a wild-type 3′UTR or mutated 3′UTR, and then were transfected with miR-324-5p mimic. The expression of Mtfr1 was assayed by immunoblot
