Molecular targets of gene therapy

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Presentation transcript:

Molecular targets of gene therapy Dipak K Das, PhD, Richard M Engelman, MD, Nilanjana Maulik, PhD, John A Rousou, MD, Joseph E Flack, MD, David W Deaton, MD The Annals of Thoracic Surgery Volume 68, Issue 5, Pages 1929-1933 (November 1999) DOI: 10.1016/S0003-4975(99)01015-2

Fig 1 Down-regulation of bcl-2 gene in the ischemic reperfused heart and its adaptive induction by repeated ischemia and reperfusion. Isolated rat hearts were made ischemic for 30 minutes followed by 2 hours of reperfusion. Experiments were terminated at baseline, after 30 minutes of ischemia and after 2 hours of reperfusion, and hearts were immediately frozen at liquid nitrogen temperture for subsequent examination of bcl-2 mRNA expression using bcl-2 cDNA probe. Another set of experiments were performed by subjecting the hearts to four cycles of repeated ischemia (5 minutes) each followed by 10 minutes of reperfusion. Hearts were similarly processed to examine bcl-2 gene expression. Total RNA was isolated from hearts and used (10 μg in each case) for Northern blot analysis with [32P]bcl-2 cDNA. The same blot was stripped and reprobed for [32P] GAPDH cDNA which served as housekeeping gene. Top panel shows the results of densitometric scanning (means ± SEM of six experiments per group); ∗p < 0.05 compared to baseline. (A) Baseline control; (B) 30-minute ischemia; (C) 30-minute ischemia followed by 2 hours of reperfusion; (D) Four cyclic episodes of 5 minutes of ischemia each followed by 10 minutes of reperfusion plus 30 minutes of ischemia and 2 hours of reperfusion. The Annals of Thoracic Surgery 1999 68, 1929-1933DOI: (10.1016/S0003-4975(99)01015-2)