Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Cardiac Overexpression of S100A6 Attenuates Cardiomyocyte Apoptosis and Reduces Infarct Size After Myocardial Ischemia‐Reperfusion

doi: 10.1161/JAHA.116.004738

Figure Lengend Snippet: Infarct size and apoptosis data post–myocardial ischemia‐reperfusion. A, Representative Evans blue–stained myocardial sections during ligation of the left anterior descending coronary artery, outlining the area at risk in nontreated controls, empty plasmid–treated controls, and human‐S100A6 plasmid–treated rats. B, Representative TTC ‐stained myocardial cryosections 1 day post‐I/R in nontreated controls, empty plasmid–treated controls, and human‐S100A6 plasmid–treated rats. The nonviable/infarcted myocardium is white, and viable myocardium is red/pink. C, Representative MTC ‐stained myocardial cryosections 28 days post‐I/R in nontreated controls, empty plasmid–treated controls, and human‐S100A6 plasmid–treated rats. The collagen debris in the central compact necrotic region of the infarcted area is shown in blue, myocardial fibers in pink, and nuclei in purple. D, Risk area by Evans blue as percentage of the LV (mean± SEM , N=6 per group). E, Nonviable myocardium/infarct area by TTC staining as percentage of the LV (mean± SEM , * P <0.05 vs nontreated and empty plasmid–treated controls, N=9 per group). F, Infarct area/scar size by MTC staining as a percentage of the LV (mean± SEM , * P <0.05 vs nontreated and empty plasmid–treated controls, N=8‐9 per group). G, IHC staining of infarct region of myocardium 1 day post‐I/R (sarcomeric α‐actinin [red], nuclei [blue], and fluorescein isothiocyanate [ FITC ]‐positive apoptotic nuclei [green]) (magnification ×40). H, Data on apoptotic index in infarct and noninfarct regions in myocardial cryosections 1 day post‐I/R in human‐S100A6 plasmid–treated and empty plasmid–treated rats compared to nontreated controls (mean± SEM , * P <0.05 vs nontreated and empty‐treated controls, N=7‐9 for each group). I, Caspase 3/7 activity in the infarct and noninfarct regions of myocardium 1 day post‐I/R in S100A6‐treated and empty plasmid–treated rats compared to nontreated controls (mean± SEM , * P <0.05 vs nontreated and empty‐treated controls, N=6 in each group). LV, left ventricle; MTC, Masson trichrome; TTC, 2,3,5‐triphenyl‐tetrazolium chloride.

Article Snippet: We also used Masson Trichrome Stain Kit (Polysciences Inc, Warminster, PA; catalog #25088‐100) to detect collagen fibers (stained blue) and necrotic tissue in the infarcted area of the LV myocardium (stained pink).

Techniques: Staining, Ligation, Plasmid Preparation, Immunohistochemistry, Activity Assay