Fig. 1. Synd1ko mice express no synd1 mRNA or protein, and their epithelial tissues appear normal. (A) The targeting vector was a MboI/SalI fragment with homology to the endogenous gene and includes 1.4 kb of the 5' flanking sequence, the entire 0.3 kb first exon and 5.3 kb of the first large intron. Positive (PGK neo) and negative (MC1-tk) selection markers were used. The disrupted allele was 8.6 kb versus the wild-type allele, which was 7.1 kb. (B) Using a 3' probe, Southern blot analyses of tail DNA isolated from two litters of mice included wildtypes (^+/+), heterozygotes (^+/-) and homozygotes (^-/-) for the disrupted gene. (C) Northern blots show that, when equal amounts of RNA isolated from wild-type and ^-/- mouse skin are run out on gels and transferred to nitrocellulose, the synd1ko mice have no detectable synd1 mRNA, whereas the wild-type mice do. Replicate blots probed with a synd4 cDNA, serving as a positive control for RNA quality, confirm the presence of synd4 mRNA in both wild-type and ^-/- tissues, but there is no upregulation in the synd1ko tissues. RNA sizes are indicated on the right; synd1 mRNA is present as two discrete bands of 2.6 and 3.4 kb, whereas synd4 mRNA is 2.6 kb. (D) Using confocal microscopy, skin and corneal tissues from wild-type and synd1ko mice were evaluated to determine the degree of colocalization of synd1 and ß4 integrin, 3 integrin and E-cadherin or 9 integrin and ZO-1. Merged images are presented. Although E-cadherin is expressed at the lateral membranes of basal cells, it is less abundant at this site, and the large amount of 3 integrin in basal cells makes it difficult to document E-cadherin—3-integrin co-expression in the basal cells in these merged images. For the 9/ZO-1 merged images of the corneal limbal region, the central cornea is located to the left and the conjunctiva is to the right. No differences in the localization of integrins, E-cadherin or ZO-1 between wild-type and synd1ko tissues are observed. Bar in D, 90 µm.

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