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J Am Coll Cardiol, 2000; 35:1769-1777
© 2000 by the American College of Cardiology Foundation
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Upregulation of the Bcl-2 family of proteins in end stage heart failure

Najma Latif, PhDa, Mahboob A. Khan, PhDa, Emma Birks, MRCPa, Aldo O’Farrell, MSca, Jules Westbrook, BSca, Michael J. Dunn, PhDa and Magdi H. Yacoub, FRCS, FACCa

a Department of Cardiothoracic Surgery, National Heart and Lung Institute, Imperial College School of Science, Technology and Medicine, Heart Science Center, Harefield Hospital, Harefield, United Kingdom



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Figure 1 (A) Top panel shows the expression of Bcl-2 in myocardial samples: donor (lanes 1–3), IHD (lanes 4–6) and DCM (lanes 6–10). The number at the left indicates MW in kD and lower band demonstrates tubulin reactivity as a marker of equal loading. Bottom panel shows the spread and the mean levels of Bcl-2 (indicated by the horizontal line) in donor, IHD and DCM. *p < 0.05. (B) Top panel shows the expression of Bcl-xL in myocardial samples: donor (lanes 1–3), IHD (lanes 4–6) and DCM (lanes 6–10). The number at the left indicates MW in kD, and lower band demonstrates tubulin reactivity as a marker of equal loading. Bottom panel shows the spread and the mean levels of Bcl-xL (indicated by the horizontal line) in donor, IHD and DCM. *p < 0.05. (C) Top panel shows the expression of Bax in myocardial samples: donor (lanes 1–3), IHD (lanes 4–6) and DCM (lanes 6–10). The number at the left indicates MW in kD, and lower band demonstrates tubulin reactivity as a marker of equal loading. Bottom panel shows the spread and the mean levels of Bax (indicated by the horizontal line) in donor, IHD and DCM. *p < 0.05. (D) Top panel shows the expression of Bak in myocardial samples: donor (lanes 1–3), IHD (lanes 4–6) and DCM (lanes 6–10). The number at the left indicates MW in kD, and lower band demonstrates tubulin reactivity as a marker of equal loading. Bottom panel shows the spread and the mean levels of Bak (indicated by the horizontal line) in donor, IHD and DCM. *p < 0.05. DCM = dilated cardiomyopathy; IHD = ischemic heart disease.

 


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Figure 2 Transverse sections of myocardial tissue showing Bcl-2 reactivity in donor (A) and in DCM (B), Bcl-xL reactivity in donor (C) and in DCM (D), Bak in donor (E) and in IHD (F) and Bax in donor (G) and in IHD (H). Original magnification x400. DCM = dilated cardiomyopathy; IHD = ischemic heart disease.

 


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Figure 3 (A) Negative TUNEL reactivity, (B) positive TUNEL DNAse 1 treated control, (C) TUNEL positive reactivity (green) in a DCM sample, (D) propidium iodide (PI) staining (red) of section C, (E) double exposure of section C and D showing a myocyte that is positive for TUNEL and for PI (orange) and (F) alpha-sarcomeric actin staining of a serial section of C. DCM = dilated cardiomyopathy; TUNEL = terminal deoxynucleotidyl transferase [TdT]-mediated dUTP nick end labelling.

 


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Figure 4 Visualization of genomic DNA extractions of myocardial tissue from donor (lanes 7 and 12), IHD (lanes 8–11) and DCM (lanes 1–6) patients using SYBR green I DNA staining. In lane 10 the beginnings of a ladder can be seen. L = 1kb DNA ladder markers; P = apoptosis positive control.

 





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