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. 2011 Sep;31(18):3832–3844. doi: 10.1128/MCB.05744-11

Fig. 4.

Fig. 4.

Generation of Aα-floxed (F5-6) and Aα knockout (Δ5-6) mice. (A) The region targeted reaches from before exon 3 past exon 11 (black horizontal line, second row, Aα wild type [WT]). The targeting construct contains (i) a loxP site (open triangle) 5′ of exon 5, (ii) a neo cassette flanked by FRT sites (black triangles) 3′ of exon 6, and (iii) a second loxP site 3′ of neo (third row). F5-6neo ES cells and mice were identified by Southern blotting of EcoRV-digested DNA (Eco) with the 3′ probe (black box) (second and fourth rows). F5-6 and Δ5-6 mice were identified by PCR using primer P>, binding to the 5′ loxP site, and primer <I, binding to intron 6 past neo (fifth and sixth rows). (B) Southern blotting of EcoRV-digested DNA with the 3′ probe identifies WT (24,000 bp) and F5-6neo (Fneo) alleles (8,200 bp). (C) PCR screening with primer pair P> and <I identifies F5-6 (F) (1,836 bp), Δ5-6 (Δ) (1,032 bp), and Δ5-6/F5-6 (1,032 and 1,836 bp) mice. (D) F5-6 and Δ5-6 mice were routinely screened with primer U> binding 5′ of the targeted region and primer <P binding to the 5′ loxP site (4,507 bp).