FIGURE 2.
Generation of Phd2f/f and Phd2+/− mouse lines. A, targeting strategy. Boxes denote exons, with numbers indicating exon number and black indicating coding sequence. neo, neomycin selection cassette; TK, thymidine kinase negative selection cassette. The sites of the PHD2rec55 (a), Pex2–3 3′ (b), Pint1 Sal 5′ (c), and Pint2–3 3′ (d) primers for PCR genotyping are as shown. Shaded boxes labeled 5′ and 3′ indicate the locations of 5′ and 3′ Southern blot probes, respectively. B and C, Southern blots of DNA from ES cells (B) or mouse tail (C). DNA was digested with ScaI (5′ Southern) or Bgl II (3′ Southern). D, PCR genotyping of mice with the indicated Phd2 genotypes. In the left panel, the WT allele produces a 0.73-kb band, whereas the floxed allele produces a 0.84-kb band. In the right panel, the 0.56-kb band indicates the presence of the knock-out allele.