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Fig. 1. Expression of COUP-TFII in the stomach using lacZ
knock-in model. (A). Generation of the lacZ knock-in allele and
generation of floxed COUP-TFII allele. Using homologous
recombination, a targeting construct containing nuclear lacZ, Neo/TK
and LoxP sites were inserted into the genomic COUP-TFII
locus, generating a targeted allele in ES cells. Treatment with Cre
recombinase and FIAU selection resulted in a lacZ knock-in allele in
which lacZ gene expression was controlled by the endogenous
COUP-TFII promoter when recombination took place between the first
and the third loxP sites of the targeted allele. In addition, floxed
COUP-TFII ES clones that retains COUP-TFII locus but lacks
selection markers were generated when recombination took place between the
second and the third loxP sites. B, BamHI; H, HindIII; S,
SalI; X, XbaI. (B) Cryostat sections of E12.5 heterozygous
COUP-TFII/lacZ knock-in embryo were stained (for 2 hours) for
lacZ activity. There is relatively high expression in the mesenchymal
cells just adjacent to the epithelium. (C). The stomach from a 3-day-old
heterozygous knock-in animal was dissected and whole-mount X-gal staining was
performed. The boundary between stomach and duodenum is indicated by
arrowhead. (D) A cryostat section of stomach from adult heterozygous knock-in
animal was stained for lacZ activity (blue) and counterstained with
propidium iodide (red). DBA lectin immunostaining denotes the parietal cells
(green). There is strong X-gal staining in the base layer and negligible
staining in the surface pit layer of the adult Zymogenic unit. m, mesenchyme;
e, epithelium; fs, fore-stomach; hs, hind-stomach; d, duodenum; oe,
esophagus.
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