Figure 2
Generation of mice with Gas2 inactivation.
(A) Targeted disruption of Gas2. The illustration includes Gas2 exon organization (empty boxes are untranslated regions (UTRs), black boxes are open reading frames), Xba1 restriction sites and the 5′ and 3′ probes used for confirmation of the mutant by southern blot, the targeting vector containing a GFP::Cre::ERT2 cassette, poly-A region, and FRT flanked kan/neo resistance, as well as the position of Gas2F/R and mutR genotyping primers. (B) Southern blot analysis of Gas2 mice with 5′ and 3′ probes showing wild type bands at 10.3 kb, 5′ mutant band at 6.8 kb, and 3′ mutant band at 6.5 kb. (C) Western blot analysis of total kidney protein showing absence of the 35 kDa Gas2 protein in the mutant mice. (D) RT-PCR analysis of mRNA expression in Gas2 −/− mice. Total RNA was extracted from kidney (1) and ovary (2) of wild-type and mutant mice. Primers amplify between exons 1–5, exons 2–4, exons 3–5, and exons 7–8 of the Gas2 cDNA. Primers against GAPDH were used as the control.
