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 3. Ube2s augments HIF-1α activation and decreases apoptosis after MI/R injury. (AC) C57BL/6 mice were intra-myocardially infected with lentivirus expressing vector control or Ube2s 48 h prior to MI/R surgery. Following 24 h of reperfusion, the protein level of HIF-1α, Bax and Bcl-2 in the heart was analyzed by Western blotting. Samples from sham group were used as controls. Each group includes 8 mice. β-Actin was used as a loading control. The representative band images (A) and relative band intensity analysis of HIF-1α (B) and ratio of Bax/Bcl-2 (C) are presented. 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 heart sections were prepared. The apoptosis was detected using TUNEL staining. The statistical analysis of percentage of TUNEL positive cells is shown. Data are mean ± SD. Data were compared using Student’s t-test. **, P < 0.01. (E) 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 the nmol AFC/h/mg protein. Data are mean ± SD. Data were compared using Student’s t-test. **, P < 0.01. (F) C57BL/6 mice were intra-myocardially transfected with control siRNA (siCtrl) or Ube2s siRNA (siUbe2s) 48 h prior to MI/R surgery. Following 24 h of reperfusion, the protein level of HIF-1α, Bax and Bcl-2 in the heart was analyzed by Western blotting. Samples from sham group were used as controls. Each group includes 8 mice. β-Actin was used as a loading control. The representative band images are presented.