Figure 3.

Generation and genotyping of floxed Des1 mice. A) Exon 2 of the Des1 gene was targeted for deletion by insertion of parallel flanking LoxP sequences. The targeting vector also contained a neomycin-resistance cassette, flanked by FLP recombinase targeting sequences, to allow for selection of successfully targeted embryonic stem cells. The neomycin cassette was subsequently removed by crossing floxed Des1 mice with FRT germline deleter mice to yield mice lacking the neomycin cassette that were used for further breeding and analysis. The green and red triangles represent LoxP and FRT sequences, respectively. PCR primer binding sites and sizes of expected products are shown on the figure. Primers sequences are shown in Supplemental Table S1. HSV-TK, herpes simplex virus thymidylate kinase gene. B) PCR genotyping results for wild-type (Des1^+/+), heterozygous (Des1^F/+), and homozygous (Des1^F/F) mice.