Reduction of cardiomyocyte apoptosis and inflammatory response by Bcl-2 overexpression during ischemia-reperfusion injury in cardiac allografts. Hearts of FVB WT or Bcl-2 Tg mice were transplanted into C57BL/6 mice. At 2 hours of reperfusion, the cardiac allografts were harvested, and histologic sections and lysates from the cardiac allografts were prepared as described in “Materials and methods.” (A) The number of apoptotic cells in cardiac allografts was counted by ISOL TUNEL staining. (B) Superoxide production was measured in excised tissue by the spin trap method. (C) Cytochrome c levels in mitochondria and cytosol were measured by ELISA, the percentage of cytochrome c release from mitochondria to cytosol was calculated, and the activities of (D) caspase-3, (E) caspase-9, and (F) caspase-8 and the levels of (G) Fas in heart lysates were measured by ELISA. (H) MPO activity was determined by MPO activity assay. (I) TNF-α, and (J) MCP-1/CCL2 production were measured by ELISA. Filled columns show the data from WT heart grafts (n = 5), and open columns show data from the Bcl-2 Tg heart grafts (n = 5). Mean ± SD values are shown. NS indicates not significant.