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Upregulation of the Bcl-2 family of proteins in end stage heart failure

^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

View larger version (35K): [in a new window]   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.

View larger version (116K): [in a new window]   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.

View larger version (65K): [in a new window]   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.

View larger version (83K): [in a new window]   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.