Figure 1.

Mtfr1 regulates mitochondrial fission and apoptosis in vitro. (a) A/R induces an increase in Mtfr1 levels. Cardiomyocytes were treated with A/R at indicated time. Mtfr1 levels were analyzed by immunoblot. (b) Knockdown of Mtfr1 attenuates the increased Mtfr1 levels upon A/R treatment. Cardiomyocytes were infected with adenoviral constructs of Mtfr1 siRNA or its scrambled form (sc) and then exposed to A/R. Mtfr1 levels were analyzed by immunoblot. (c and d) Knockdown of Mtfr1 attenuates mitochondrial fission induced by A/R treatment. Cardiomyocytes were infected with adenoviruses harboring Mtfr1 siRNA or its sc form and then treated with A/R. Mitochondria were stained by MitoTracker red and the nuclei were visualized by DAPI. (c) Scale bar, 20 μm (upper panel); scale bar, 5 μm (lower panel). The cells with fragmented mitochondria were counted. (d) *P<0.05 versus A/R alone. (e and f) Knockdown of Mtfr1 reduces A/R-induced apoptosis. Cardiomyocytes were infected with adenoviruses harboring Mtfr1 siRNA or its sc form and then were subjected to 2-h anoxia followed by 12-h reoxygenation. TUNEL was employed to analyze apoptotic cells. TUNEL-positive cells were counted and calculated. (e) The caspase-3 activity was analyzed by using an Apo-ONE Homogeneous Caspase-3/7 assay kit (Madison, WI, USA) (f) *P<0.05 versus A/R alone