Figure 2: miR-421 participates in the regulation of Pink1 expression. | Nature Communications

Figure 2: miR-421 participates in the regulation of Pink1 expression.

From: E2F1-dependent miR-421 regulates mitochondrial fragmentation and myocardial infarction by targeting Pink1

Figure 2

(a) MiR-421 levels were analysed by qRT–PCR. Data are shown as mean±s.e.m. of four independent experiments. *P<0.05 versus untreated group in Student’s t-test. (b) Putative miR-421-binding site in the 3′-UTR region of Pink1. (c) Cardiomyocytes were infected with adenoviral miR-421 or β-gal. The levels of Pink1 were analysed by immunoblot. Data are shown as mean±s.e.m. of three independent experiments. *P<0.05 versus untreated group in Student’s t-test. (d) Cardiomyocytes were transfected with miR-421 antagomir (anta-421) or the antagomir negative control (anta-NC), then exposed to H2O2. The levels of Pink1 were analysed by immunoblot. Data are shown as mean±s.e.m. of three independent experiments. Analysis was performed with one-way analysis of variance (ANOVA) followed by Tukey–Kramer post hoc analysis. *P<0.05. (e) Pink1 target protector attenuates the reduction of Pink1 induced by miR-421. Cardiomyocytes were infected with adenoviral miR-421, then transfected with the target protector (Pink1-TPmiR-421) or the control (Pink1-TPcontrol). Pink1 levels were detected by immunoblot. Data are shown as mean±s.e.m. of three independent experiments. Analysis was performed with one-way ANOVA followed by Tukey–Kramer post hoc analysis. *P<0.05. (f) Pink1 target protector inhibits H2O2-induced Pink1 downregulation. Cardiomyocytes were transfected with the Pink1-TPmiR-421 or Pink1-TPcontrol, and then exposed to H2O2. The levels of Pink1 were analysed by immunoblot. Data are shown as mean±s.e.m. of three independent experiments. Analysis was performed with one-way ANOVA followed by Tukey–Kramer post hoc analysis. *P<0.05. (g) Pink1 wild-type (WT) 3′-UTR and a mutated 3′-UTR in the miR-421-binding site are shown. (h) miR-421 suppresses Pink1 translation. HEK293 cells were infected with adenoviral miR-421 or β-gal, then transfected with the luciferase constructs of the WT Pink1-3′-UTR (Pink1-3′-UTR-WT) or a mutated Pink1-3′-UTR (Pink1-3′-UTR-mut). The luciferase activity was analysed. Data are shown as mean±s.e.m. of four independent experiments. Analysis was performed with one-way ANOVA followed by Tukey–Kramer post hoc analysis. *P<0.05.

Back to article page