Supplemental Figure 1
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Fig.
S1. Targeting of
the Ext2 gene.
(A) Schematic diagram showing the structure of the Ext2 wild-type locus, the
targeting vector and the mutant allele. The top line shows a partial
restriction map of the Ext2 locus. The Ext2 gene consists of 14 exons, the second exon
(E2) contains the translation initiation codon. The targeting vector contains a
PGK-neo cassette in the opposite transcriptional orientation to the Ext2 and a
PGK-tk cassette. The regions between the broken lines represent the 5¢ and 3¢ regions of homology. Positions of the
fragments used as the 5¢ and 3¢ external
probes in Southern blotting are shown at the top. The sizes of the predicted
fragments for the wild-type and mutant alleles in Southern blot analyses are
shown on the bottom. Arrows indicate the positions of the primers used for PCR
genotyping. B, BsteII;
H, HindIII; R, EcoRI; S, SpeI; Sm, SmaI. (B) Southern blot analyses of
genomic DNA from E6.5 embryos generated from matings between Ext2 heterozygous
mice. DNA was digested with HindIII and hybridized with probe B. (C) PCR amplification of genomic DNA
from E6.5 embryos using a four-primer PCR strategy shows a 190 bp wild-type
fragment and a 260 bp mutant fragment.
