A mechanistic role for cardiac myocyte apoptosis in heart failure
J. Clin. Invest. Detlef Wencker, et al. 111:1497 doi:10.1172/JCI17664 [Go to this article.]

Figure 1
Expression of a conditionally active caspase-8 allele in the hearts of transgenic mice. (a) Structure of the transgene protein and strategy for its activation. M, myristoylation signal; FKBP, human FKBP-12 (pk mutant); p20 and p10, 20- and 10-kDa domains, respectively, of human procaspase-8; HA, hemagglutinin tag; FK1012, dimerizer (see text). Cysteine 360, a residue essential for caspase activity, is shown. (b) Southern blot analysis of EcoRI-digested genomic DNA from WT mice and two of the ten transgenic lines generated. The probe, an EcoRI-XbaI mouse α-cardiac myosin heavy chain genomic fragment, identified a 3.6-kb transgene fragment and a 2.5-kb fragment of the endogenous α-cardiac myosin heavy chain. (c) Immunoblot analysis using an antibody against human caspase-8, showing transgene protein expression in the hearts of the most highly expressing (line 7) and least highly expressing (line 169) lines at 3 weeks of age. Levels of the point-mutated transgene protein in the hearts of line C360A mice are similar to those of the catalytically active transgene protein in the hearts of line 7 mice. The transgene protein was not detectable in a survey of other organs, as expected with the cardiac myocyte–specific α-cardiac myosin heavy chain promoter (not shown). The lower portion of the blot was reacted with an antibody against mouse tubulin as a loading control.