Figure EV1. Generation of Vegfr2 ^{Y1212F/Y1212F} mice.

• A
  Mice were generated using site‐directed mutagenesis on a genomic fragment of the mouse Vegfr2/Flk1 gene. A substitution mutation, TAT to TTC, was introduced into exon 27 and a mutation‐containing targeting vector (red) was transfected into embryonic stem cells, where the vector was inserted into the wild‐type (WT) sequence (blue) by homologous recombination. The neo‐cassette was removed by crossing with a CAG‐Cre‐recombinase expressing mouse line, resulting in an 86‐base pair remaining insertion into intron 26.
• B
  The correct introduction of the mutation and the otherwise unaffected sequence of the region surrounding exon 27 were verified by Sanger sequencing of forward and reverse strands of DNA from Vegfr2 ^{Y1212F/Y1212F} mice, extracted from ear biopsies. The WT and Vegfr2 ^{Y1212F/Y1212F} mutant sequence were aligned. The 1212 codon (blue, tyrosine, wild‐type; red, phenylalanine, mutant) and the 86‐base pair insertion (yellow) are indicated.