Research Paper Volume 12, Issue 7 pp 5716—5732

Ube2s-stabilized β-catenin protects against myocardial ischemia/reperfusion injury by activating HIF-1α signaling

Figure 6. β-Catenin knockdown abrogates Ube2s-augmented HIF-1α activation and diminishes Ube2s protective effect on MI/R injury. (AB) C57BL/6 mice were intra-myocardially infected with lentivirus expressing vector control or Ube2s in combination with the transfection with control siRNA (siCtrl) or β-Catenin siRNA (siβ-Catenin) 48 h prior to MI/R surgery. Following 24 h of reperfusion, the protein expression of targets as indicated in the heart was analyzed by Western blotting. Each group includes 8 mice. β-Actin was used as a loading control. The representative band images (A) and relative band intensity analysis (B) are presented. Data are mean ± SD. Data were compared using Student’s t-test. **, P < 0.01; NS, not significant. (C) Heart samples were harvested as described in (A). The supernatants of the homogenized heart samples were collected, and the caspase-3 activity was determined. The results are expressed as nmol AFC/h/mg protein. Data are mean ± SD. Data were compared using Student’s t-test. **, P < 0.01. (D) Heart samples were harvested as described in (A), and the mid-myocardial cross sections were prepared. The infarct size in the heart sections was quantified, and the results of percentage of size are shown. AAR/LVA, ratio of area at risk (AAR) to left ventricular area (LVA); IA/AAR, ratio of infarct area (IA) to AAR. Data are mean ± SD. Data were compared using Student’s t-test. **, P < 0.01; *, P < 0.01; NS, not significant. (E) C57BL/6 mice were treated as in (A). The samples of serum were collected and the level of creatine phosphokinase (CPK) was quantified. Data are mean ± SD. Data were compared using Student’s t-test. **, P < 0.01.