Fig. 1.

Generation of Cldn11/Cldn14 double knockout mice. a Shown are the Cldn14 and Cldn11 genotypes of mice used in each set of matings and the genotypes of the offspring. The numbers above the genotypes indicate the proportion of mice with this genotype among all offspring in a given mating. b For specific amplification of the wt and the knockout (ko) alleles, if present, from each DNA sample, three different PCR-primers were used in each case (a common primer, a wt primer, and a mutant primer). For Cldn11 a single multiplexed PCR reaction with all three different PCR-primers is performed. PCR product sizes are 390 bp for the wt allele and 290 bp for the ko allele. For Cldn14 two separate PCR reactions with two sets of primers (common and wt primers or common and mutant primers) are performed. PCR product sizes are 340 bp for the wt allele and 275 bp for the ko allele. c Brain cDNA was analyzed by RT-PCR for Cldn11 and Cldn14 expression (512 and 664 bp products, respectively) in mice of various genotypes. As a control, the Actb gene (β-actin) (465 bp product) was equally amplified in all samples.