Fig. 2.

(NADPH) oxidase homolog 1 (NOH-1) is a target for miR-145-5p. To explore the interaction between miR-145-5p and NOH-1, HEK-293T cells were co-transfected with NOH-1 3’ untranslated region (UTR) (wild- and mutant-type) and miR-145-5p mimics or negative control (NC) mimics. Then, H9c2 cells were transfected with miR-145-5p mimics, NC mimics, miR-145-5p inhibitors, or NC inhibitors. (A) A schematic diagram is shown for the predicted miR-145-5p binding sites within the 3’ UTR of NOH-1. Sequences of candidate binding sites between miR-145-5p and NOH-1 are marked in red, while sequences with the point mutation in the target candidate sites are in green. (B) A luciferase reporter assay was performed to measure the luciferase activity. (C) qRT-PCR was performed to detect the expression of miR-145-5p. (D, E) qRT-PCR and western blot analyses were used to detect the NOH-1 expression at the mRNA and protein levels. All results are presented as means ± SD from three independent experiments, and they were analyzed by one-way ANOVA. ns, not significant. ^$$$P<0.001, compared with NC mimics + NOH-1 3’UTR WT. ^aP<0.05, compared with miR-145-5p mimics + NOH-1 3’UTR WT. ⁺⁺P<0.01, compared with NC mimics. ⁺⁺⁺⁺P<0.0001, compared with NC mimics. ^{^^}P<0.01, compared with NC inhibitors. ^{^^^^}P<0.0001, compared with NC inhibitors.