Figure 1: Pink1 inhibits mitochondrial fragmentation and apoptosis.

(a) Cardiomyocytes were treated with 150 μM H₂O₂ at indicated time. Pink1 levels 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. (b) Cardiomyocytes were infected with adenoviral Pink1 and β-gal, then exposed to H₂O₂. Pink1 levels 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. (c) Cardiomyocytes were treated as described in b. The cells with fragmented mitochondria were counted (right panel, upper panel). Bar=20 μm. TdT-mediated dUTP nick end labelling (TUNEL) was employed to analyse apoptotic cells (right panel, lower panel). 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 versus H₂O₂ alone. (d) WT and Pink1-transgenic mice were subjected to I/R as described in Methods. Pink1 levels were detected by immunoblot. 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. (e,f) Pink1-transgenic mice represent attenuated mitochondrial fragmentation and apoptosis on I/R injury. The fragmented mitochondria were shown in e. TUNEL assay was performed to detect apoptotic cells (f). Data are shown as mean±s.e.m. of six independent experiments. Analysis was performed with one-way ANOVA followed by Tukey–Kramer post hoc analysis. *P<0.05 versus WT+I/R. (g) Pink1-transgenic mice show attenuated myocardial infarct sizes in response to I/R. The upper panels are representative photos of midventricular myocardial slices. The lower panel shows infarct sizes. LV, left ventricle; INF, infarct area. Data are shown as mean±s.e.m. of six independent experiments. Analysis was performed with one-way ANOVA followed by Tukey–Kramer post hoc analysis. *P<0.05 versus WT+I/R. Bar=2mm. (h) Pink1-transgenic mice present markedly preserved cardiac function after I/R. Transthoracic echocardiographic analysis was performed. Data are shown as mean±s.e.m. of six independent experiments. Analysis was performed with one-way ANOVA followed by Tukey–Kramer post hoc analysis. *P<0.05.