1. Date of download: 5/28/2016 Copyright © The American College of Cardiology. All rights reserved. From: Apoptotic pathway activation from mitochondria and death receptors without caspase-3 cleavage in failing human myocardium: Fragile balance of myocyte survival? J Am Coll Cardiol. 2002;39(3):481-488. doi:10.1016/S0735-1097(01)01769-7 Comparison of left ventricular procaspase-9, activated caspase-9 and caspase-9S expression in patients with end-stage heart failure with nonfailing donor hearts. The messenger ribonucleic acid (mRNA) expression of caspase-9S was downregulated and the ratio of caspase-9S/procaspase-9 was decreased in patients with heart failure (a). Western blot analyses of procaspase-9, activated caspase-9 and caspase-9S protein in failing left ventricles, as compared with donor ventricles (b), illustrate increases of procaspase-9 by 25% and activated caspase-9 by 65% and an inverse relationship for caspase-9S, with no detection of caspase-9S in the failing myocardium (c). ( ∗ )p < 0.1; ∗∗ p < 0.01; ∗∗∗ p < 0.001. Figure Legend:

2. Date of download: 5/28/2016 Copyright © The American College of Cardiology. All rights reserved. From: Apoptotic pathway activation from mitochondria and death receptors without caspase-3 cleavage in failing human myocardium: Fragile balance of myocyte survival? J Am Coll Cardiol. 2002;39(3):481-488. doi:10.1016/S0735-1097(01)01769-7 Immunoblot analysis of cytochrome c and manganese superoxide dismutase (MnSOD; as a marker of the mitochondrial matrix) in the cytosol (a) and total cell lysate (b) of left ventricular specimens from donor hearts and failing hearts. In the failing myocardium, cytosolic cytochrome c was 53% higher (c), whereas MnSOD was not different between the two groups in terms of the cytosolic fraction and total lysate. In cultured rat cardiomyocytes, cytochrome c release could only be detected after treatment with 10 μmol/l of norepinephrine for 24 h (d). ∗ p < 0.05. Figure Legend:

3. Date of download: 5/28/2016 Copyright © The American College of Cardiology. All rights reserved. From: Apoptotic pathway activation from mitochondria and death receptors without caspase-3 cleavage in failing human myocardium: Fragile balance of myocyte survival? J Am Coll Cardiol. 2002;39(3):481-488. doi:10.1016/S0735-1097(01)01769-7 Analysis of left ventricular protein expression of procaspase-8, activated caspase-8, flice-like inhibitory protein (FLIP) S and FLIP L in the failing human myocardium, as compared with donor organs. The messenger ribonucleic acid (mRNA) expression of FLIP S and FLIP L is significantly downregulated in the failing myocardium (a). For activated caspase-8, there was an increase of 62% in the failing myocardium, as compared with donor hearts (b), whereas procaspase-8 was not different between the two groups. The FLIP S protein decreased significantly in the failing myocardium (b). ( ∗ )p < 0.1; ∗ p < 0.05; ∗∗∗ p < 0.001. Figure Legend:

4. Date of download: 5/28/2016 Copyright © The American College of Cardiology. All rights reserved. From: Apoptotic pathway activation from mitochondria and death receptors without caspase-3 cleavage in failing human myocardium: Fragile balance of myocyte survival? J Am Coll Cardiol. 2002;39(3):481-488. doi:10.1016/S0735-1097(01)01769-7 Comparison of left ventricular expression of the inhibitor of apoptosis proteins, hIAP-1, hIAP-2 and XIAP, from patients with end- stage heart failure and donors. The messenger ribonucleic acid (mRNA) expression of hIAP-1, hIAP-2 and XIAP was downregulated in the left ventricles of patients with end-stage cardiomyopathy (a). Immunoblot analysis showed a decrease of hIAP-1 and XIAP in patients with terminal heart failure, as compared with donor organs (b). ( ∗ )p < 0.1; ∗ p < 0.05; ∗∗∗ p < 0.001. Figure Legend:

5. Date of download: 5/28/2016 Copyright © The American College of Cardiology. All rights reserved. From: Apoptotic pathway activation from mitochondria and death receptors without caspase-3 cleavage in failing human myocardium: Fragile balance of myocyte survival? J Am Coll Cardiol. 2002;39(3):481-488. doi:10.1016/S0735-1097(01)01769-7 Immunoblot analysis of caspase-3 and its cleavage substrates gelsolin and fodrin in the left ventricles of patients with end-stage heart failure, as compared with donor hearts: 17- and 19-kDa–cleaved caspase-3 was exclusively detectable in cell lysates from cytochrome c–treated Jurkat and NIH 3T3 cell extracts. Neither donor hearts nor terminally failing hearts revealed caspase-3 activation when using an anti-active caspase-3 antibody (Cell Signaling Technology) (a). When performing an immunoblot with an antibody against the 32-kDa inactive and active caspase-3 proteins (Pharmingen), there was no cleavage product detectable, although procaspase-3 was detectable in a similar amount (b). For gelsolin, we found only the uncleaved form in both groups (c). The terminally failing hearts revealed significantly more cleaved (p < 0.001) and uncleaved (p < 0.05) fodrin, as compared with donor hearts (d). Equal protein loading of each patient has been shown by coomassie staining (e). Figure Legend:

6. Date of download: 5/28/2016 Copyright © The American College of Cardiology. All rights reserved. From: Apoptotic pathway activation from mitochondria and death receptors without caspase-3 cleavage in failing human myocardium: Fragile balance of myocyte survival? J Am Coll Cardiol. 2002;39(3):481-488. doi:10.1016/S0735-1097(01)01769-7 Messenger ribonucleic acid (mRNA) expression of pro-atrial natriuretic peptide (ANP) as an overload indicator, several apoptosis inhibitors, apoptosis-inducing caspases and mitochondrially localized proteins in the left ventricles of terminally failing hearts treated with or without beta-blockers. Values are given in optical density units normalized to donor organ values. ∗ p < 0.05. ∗∗ p < 0.01. ∗∗∗ p < 0.001. AIF = apoptosis-inducing factor; FLIP = flice-like inhibitory protein. Figure Legend: