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. 2012 Nov 19;209(12):2263–2276. doi: 10.1084/jem.20121505

Figure 3.

Figure 3.

Generation of IL-7Rα-cKO mice. (A) Targeting strategy to generate IL-7Rα-cKO allele. Numbered boxes = exons, gray boxes = out of frame exons as a result of Cre-mediated deletion. NeoR = neomycin resistance cassette. Black triangles = loxP sites. Gray ovals = frt sites. H, V = HindIII and EcoRV restriction sites, respectively. Location of 5′ and 3′ probes for Southern blot is indicated. Configuration of the IL-7Rα germline KO allele used in this study is shown for comparison sake. (B) Southern blot confirmation of proper integration of 5′ and 3′ ends of targeting construct in ES cells. HindIII restriction digest results in an 8 Kb WT fragment and a 4.7 Kb targeted fragment detected with the 5′ probe. EcoRV restriction digest results in a 14 kB WT fragment and a 11 kB targeted fragment detected with the 3′ probe as depicted in A. (C) Confirmation that after Actin-flp recombinase-mediated deletion of the NeoR cassette, expression of the IL-7Rαfl allele is equal to WT IL-7Rα expression in the absence of Cre throughout T cell development.