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Fig. 1. Generation of the targeting construct and Fgfr2^+/S252W mice. (A) Structure of a portion of the wild-type Fgfr2 gene with exons IIIa, IIIb, IIIc and 10; the targeting construct with the TK cassette and neo cassette with flanking loxP sequences; and the mutant allele produced by homologous recombination and with neo deletion mediated by Cre. The 755_756CA>GG, S252W mutation (*) was introduced into exon IIIa. Probes (–) and restriction enzymes (St, StyI; H, HindIII; S, SacI) used for Southern blot analysis, and PCR primers (F, Fn, R; arrowheads) used for genotyping are shown. (B) Identification of mutant and wild-type alleles in two independent ES cell clones using Southern blot analysis with 5' and 3' probes. (C) Genotype of a litter from +/S252W^floxx+/Cre by PCR of tail DNA. Lanes 1, 3 and 5: heterozygote with +/S252W^flox, showing wild-type allele (521 bp) and hypomorphic allele with neo (1.1 kb). Lanes 2 and 4: heterozygote mutant +/S252W, showing mutant allele after neo deletion (581 bp). Lane 7: hypomorphic mutant +/S252W^flox/S252W. Lanes 6 and 8: wild type (+/+). (D) RT-PCR detection of mutant transcript in lung and skull tissues. Other tissues tested expressed the mutant allele (data not shown). The mutation introduces an additional SfiI restriction site. (E) Gross appearance of +/S252W and wild-type postnatal day (P)1 mice. Note the small body size of the mutant mouse. The difference of body weight between the mutant and control mice was significant.

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