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Figure 6 (A) Genetic linkage of coronary artery disease (CAD)/myocardial infarction to chromosome 15q26 (adCAD1). Pedigree structure and genotypic analysis of kindred QW1576. Individuals with CAD are indicated by closed squares (men) or closed circles (women). Unaffected individuals are indicated by open symbols. Normal men under the age of 50 years or normal women less than 55 years are shown with light gray color as uncertain phenotype. Deceased individuals are indicated by a slash, "/." The proband is indicated by an arrow. Genotypes for markers D15S104, D15S212, D15S120, and D15S87 are shown below each symbol. Initial linage was identified with D15S120, which yielded a logarithm of the odds ratio score of 4.19 at a recombination fraction of 0. Haplotype cosegregating with the disease is indicated by a black vertical bar. (B) DNA sequence analysis of the wild-type (WT) allele and the 21-base pair (bp) deletion allele (21bp) of MEF2A. Sequence analysis of exon 11 of MEF2A in the proband (II.1) revealed the presence of a deletion. The WT and deletion alleles were separated by a 3% agarose gel and a single-strand conformation polymorphism (SSCP) gel, purified and sequenced directly. The location of 21bp is indicated. (C) 21bp results in a deletion of seven amino acids of MEF2A (Q₄₄₀P₄₄₁P₄₄₂Q₄₄₃P₄₄₄Q₄₄₅P₄₄₆ or 7aa). (D) Functional characterization of WT and 7aa MEF2A proteins by transcriptional activation assays. The effect of 7aa on transcription activation activity of MEF2A was analyzed in the presence or absence of the zinc-finger transcription factor GATA-1 using the ANF_–700 promoter. Transcriptional activity is shown as relative luciferase activity on the y axis. The transcriptional activity of the reporter gene only (vector) was set arbitrarily to 1. Western blot analysis with antiMEF2A rabbit polyclonal antiserum showed that both WT and mutant MEF2A (7aa) were successfully expressed in transfected HeLa cells (shown in the box; C, vector only as negative control; the MEF2A antibody detected two bands as previously reported). The data shown were from two independent experiments in triplicate, and are expressed as mean ± S.E. WT = wild type MEF2A; WT/7aa = coexpression of both wild type and mutant MEF2As; 7aa = the 7 amino acid deletion of MEF2A. (With permission from Wang et al. [36]).

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