Research Paper Volume 11, Issue 11 pp 3536—3550

Figure 5. Inhibition of IKKε/STAT-1 axis abrogates the promoted MI/R injury by TRIM6 overexpression. (A–B) The mouse heart was pre-infected in vivo with lentivirus expressing vector control (LV-vector) or Trim6 (LV-Trim6) in the presence or absence of CAY10576 or fludarabine 48 hrs before surgery. Mice were then subjected to sham surgery or experimental MI/R. Each group contained 7 mice. At 24 hrs after reperfusion, the hearts were harvested for analyses. (A) The mid-myocardial cross sections of heart were stained with TTC, and the quantification of infarct size in each group (% of LV) is shown. (B) The level of serum creatine phosphokinase (CPK) from each group was measured. (C–F) The mouse heart was pre-infected in vivo with lentivirus expressing vector control (LV-vector), wild-type Trim6 (LV-Trim6-wt) or C15A RING mutant Trim6 (LV-Trim6-mut) 48 hrs before surgery. Mice were then subjected to sham surgery or experimental MI/R. Each group contained 7 mice. At 24 hrs after reperfusion, the hearts were harvested for analyses. (C) The lysates of heart tissues were co-immunoprecipitated (co-IP) by IKKε antibody. The expression of ubiquitin (K48) in the co-IP products and input samples was measured by Western blotting analysis. (D) The protein expression of TRIM6, p-IKKε, IKKε, p-STAT1, STAT1, Bax and Bcl-2 in the heart was determined by Western blotting analysis. β-Actin was used a loading control. (E) The mid-myocardial cross sections of heart were stained with TTC, and the quantification of infarct size in each group (% of LV) is shown. (F) The level of serum creatine phosphokinase (CPK) from each group was measured. The All data are expressed as mean ± SD (n = 7). **, P < 0.01; NS, not significant.