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Figure 5 Immunoblot analysis of caspase-3 and its cleavage substrates gelsolin and fodrin in the left ventricles of patients with end-stage heart failure, as compared with donor hearts: 17- and 19-kDa–cleaved caspase-3 was exclusively detectable in cell lysates from cytochrome c–treated Jurkat and NIH 3T3 cell extracts. Neither donor hearts nor terminally failing hearts revealed caspase-3 activation when using an anti-active caspase-3 antibody (Cell Signaling Technology) (a). When performing an immunoblot with an antibody against the 32-kDa inactive and active caspase-3 proteins (Pharmingen), there was no cleavage product detectable, although procaspase-3 was detectable in a similar amount (b). For gelsolin, we found only the uncleaved form in both groups (c). The terminally failing hearts revealed significantly more cleaved (p < 0.001) and uncleaved (p < 0.05) fodrin, as compared with donor hearts (d). Equal protein loading of each patient has been shown by coomassie staining (e).
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