Fig. 1.

Generation of the tissue specific knockout of the α₂-isoform of the Na,K-ATPase. A: Cre/LoxP targeting strategy. i: Restriction map depicting the endogenous locus of the wild-type α₂ allele. ii: Targeting construct for the α₂ allele with the 5′-loxP (black triangles) and 3′-neomycin (Neo) and a thymidine kinase cassette (TK) downstream. iii: Schematic of the targeted α₂-allele following successful homologous recombination with the targeting construct. iv: Schematic of the targeted α₂ allele following transient Cre expression. v: Schematic of the deleted α₂-allele following the tissue specific expression of the SM22α Cre. B: Southern blot analysis of the 5′-loxP site following BglII restriction digests. C: Southern blot analysis of the 3′-loxP site following SpeI restriction digests. D: Genotyping of DNA isolated from tail biopsies. E: Genotyping of DNA isolated from aortas to screen for recombination between the 5′ and 3′ loxP sites. ES, embryonic stem cells.