*2.10. Infarction Size and Leukocytes Infiltration Area Analysis*

Randomly chosen histological heart sections of four horizontal infarct levels were stained with Fast Green FCF (Sigma-Aldrich, Saint Louis, Missouri, USA) and Sirius Red (Chroma Waldeck GmbH & Co. KG, Münster, Germany) assessing tissue localization and distribution of connective fibers. Two contiguous levels of the heart (*n* = 6 for each group) which represent the major infarction ratio were quantitatively estimated using computer aided image analysis (AxioVision LE Rel.4.5 software, Carl Zeiss AG, Oberkochen, Germany). To evaluate leukocytes infiltration area 48 h after the second ligation, the two contiguous levels of the heart which represent the major infarct ratio were stained with Hematoxylin (Merck, Darmstadt, Germany) and Eosin (Thermo Shandon Ltd., Runcorn, UK) and representative images were taken using computerized planimetry (AxioVision LE Rel.4.5 software).

#### *2.11. Statistical Analysis*

Statistical analysis was performed using SigmaStat (Version 3.5, Systat Software, San Jose, CA, USA) and IBM SPSS Statistics for Windows (Version 22.0., IBM Corp., Armonk, NY, USA). The comparisons of two experimental groups were performed using Mann–Whitney *U* test. *p* values ≤ 0.05 were considered as statistically significant.

#### **3. Results**
