1. αB-Crystallin Improves Murine Cardiac Function and Attenuates Apoptosis in Human Endothelial Cells Exposed to Ischemia-Reperfusion 
Jeffrey B. Velotta, MD, Naoyuki Kimura, MD, Stephanie H. Chang, BS, Jaehoon Chung, MD, Satoshi Itoh, MD, Jonathan Rothbard, PhD, Philip C. Yang, MD, Lawrence Steinman, MD, Robert C. Robbins, MD, Michael P. Fischbein, MD, PhD 
The Annals of Thoracic Surgery 
Volume 91, Issue 6, Pages (June 2011)
DOI: /j.athoracsur
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

2. Fig 1
The C57BL/6 mice underwent temporary left anterior descending artery (LAD) occlusion for 30 minutes. Phosphate-buffered saline (PBS) or αB-crystallin (CryAB) was administered intramyocardially 15 minutes before reperfusion of the LAD (PBS 100 μL; CryAB 100 μL, 50 μg) and after ischemia-reperfusion (I/R) injury (PBS 100 μL; CryAB 100 μL, 50 μg) every other day, by intraperitoneal injection. Left ventricular ejection fraction (LVEF) was measured by magnetic resonance imaging (%) of PBS-treated and CryAB-treated C57BL/6 mice 40 days after I/R injury (n = 6 per group; *p < 0.005). Data are mean ± SD.
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

3. Fig 2
Human microvascular endothelial cells (HMEC-1) were incubated with 50 μg αB-crystallin (CryAB), 500 μg/10 mL, or phosphate-buffered saline (PBS) in a hypoxia chamber for 24 hours and then allowed to reoxygenate at room air for 30 minutes. (A) Representative Western blots of PBS-treated or CryAB-treated HMEC-1 cells. (B) Enzyme-linked immunosorbent assay cytoplasmic histone-associated DNA fragment apoptosis assay of PBS-treated or CryAB-treated HMEC-1 cells (†p < 0.01). (C) Caspase-3 activity assay of PBS-treated or CryAB-treated (50 μg) HMEC-1 cells (*p < 0.005). (OD = optical densitometry.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

4. Fig 3
Densitometric analysis of Western blot in human microvascular endothelial cells (HMEC-1), in vitro. Quantitative densitometric analysis of Western blots were performed in HMEC-1 treated with αB-crystallin (CryAB) 10 μg (light gray bars) or 50 μg (dark gray bars) or phosphate-buffered saline (PBS [black bars]) after different hypoxia times (6 hours, 12 hours, and 24 hours) and then allowed to reoxygenate at room air for 30 minutes. Protein was normalized with β-actin signal. (Each treatment group, n = 3 to 4.) (A) Bcl-2; (B) free bax; (C) cleaved caspase-3; (D) cleaved caspase-9; (E) cleaved-PARP. (NS = not significant.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions

5. Fig 4
Mouse HL-1 murine atrial cardiomyocytes were incubated with 50 μg αB-crystallin (CryAB) 500 μg/10 mL, or phosphate-buffered saline (PBS) in a hypoxia chamber (1% O2, 5% CO2, 94% N2) for 24 hours and then allowed to reoxygenate at room air for 30 minutes. (A) Representative Western blots of PBS-treated or CryAB-treated HL-1 cells. (B) Enzyme-linked immunosorbent assay cytoplasmic histone-associated DNA fragment apoptosis assay of PBS-treated or CryAB-treated HL-1 cells. (C) Caspase-3 activity assay of PBS-treated or CryAB-treated HL-1 cells. (OD = optical densitometry.)
The Annals of Thoracic Surgery  , DOI: ( /j.athoracsur )
Copyright © 2011 The Society of Thoracic Surgeons Terms and Conditions