Fig. 1.
Generation of a conditional Hif-2α allele. (A) Schematic of the targeting vector and recombination steps. (B) PCR genotyping of mice with Cre activated as young pups. Lanes 1 –3 show Hif-2αfl/fl mice without Cre expression. Lane 4* shows a Hif-2αfl/Δ animal without Cre expression. Samples in lanes 5–7 represent DNA from mice which harbor the Cre transgene and show loss of Hif-2α. (C) Southern blot analysis of DNA isolated from the kidney (lanes 1, 4, 7), the liver (lanes 2, 5, 8), and the lung (lanes 3, 6, 9) of Cre negative and positive animals. (D) RT-PCR analysis of cDNA from kidneys and livers of six different animals. Lanes 1 and 7 show floxed mRNA transcripts still containing exon 2 (650 bp). Lanes 2*, 3*, 8*, and 9* express both the floxed and deleted exon 2 mRNA, and a background band. Depletion of Hif-2α results in total loss of exon 2 as displayed in lanes 4–6, and 10–12 (450 bp). (E) β-Galactosidase expression after Cre activation in ROSA26 reporter mice.