Fig. 1. Targeted disruption of the nectin 1 and nectin 3 genes. (A) Targeted disruption of the nectin 1 gene. (A) (a) The structure of the mouse nectin 1 gene with coding exons 2-5 is shown at the top. A targeting vector was designed to remove the exon 2. The construct contained 2.8 kb 5' flanking sequence and 4.0 kb 3' flanking sequence. (b) Southern blotting of ES clones. HindIII-digested DNAs derived from ES cells were hybridized with the 5'- or 3'-probe. WT, wild-type; +/–, nectin 1^+/–. (c) Genotyping by PCR of genomic DNA extracted from littermate mice at 21 days of age. Wild-type (WT) and targeted alleles give the bands of 639 bp and 459 bp, respectively. +/–, nectin 1^+/–; –/–, nectin 1^–/–. (d) Western blotting of the extracts from the brains of wild-type and nectin 1^–/– mice with the anti-nectin 1 and anti-actin antibodies. (B) Targeted disruption of the nectin 3 gene. (a) The structure of the mouse nectin 3 gene with coding exon 1 is shown at the top. A targeting vector was designed to remove the exon 1. The construct contained 6.5 kb 5' flanking sequence and 4.2 kb 3' flanking sequence. (b) Southern blotting of ES clones. BamHI-digested DNAs derived from ES cells were hybridized with the 5'- or 3'-probe. WT, wild-type; +/–, nectin 3^+/–. (c) Genotyping by PCR of genomic DNA extracted from littermate mice at 21 days of age. Wild-type and targeted alleles give the bands of 379 bp and 814 bp, respectively. WT, wild-type; +/–, nectin 3^+/–; –/–, nectin 3^–/–. (d) Western blotting of the extracts from the brains of wild-type and nectin 3^–/– mice with the anti-nectin 3 and anti-actin antibodies.