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. 2008 Feb 1;118(2):651–658. doi: 10.1172/JCI32899

Figure 1. Eya4-targeting strategy.

Figure 1

(A) A Neo and Zeo cassette flanked by loxP sites (black triangles) was inserted into Eya4 exons 8–10 by homologous recombination in bacteria. The mutant allele was introduced into ES cells using standard homologous recombination techniques (see Methods). (B) PCR analyses of wild-type (+/+), homozygous Eya4–/– (–/–), and heterozygous Eya4+/– (+/–) genotypes. Primers intron 7F (I7F), exon 8R (E8R), and NeoR demonstrated the presence of wild-type (516 bp) and mutant (446 bp) alleles. (C) RT-PCR of cardiac cDNA using external forward primer 4F and internal reverse primer 9R identified a single 469-bp product in wild-type (+/+) and heterozygous Eya4+/– (+/–) but not Eya4–/– (–/–) mice.