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Fig. 8. ß-catenin is required for the expression of Sox17 targets genes at
gastrula stage. (A) Two-cell embryos were vegetally injected with either 20 ng
of antisense ß-catenin morpholino oligos (MO) and/or 100 pg of RNA
encoding a N-terminal deleted form of stabilized ß-catenin
(
N-ß-cat) (Yost et al.,
1996), which does not contain the sequence targeted by the
antisense oligo. At a series of stages throughout gastrulation (stages 10, 11
and 12) whole embryos were harvested and assayed by real-time RT-PCR for the
expression of Sox17 target genes as well as several other control genes. The
histograms show the relative expression levels normalized to the loading
control, ODC. For simplicity only the stage 11 data (stage 10 for
Xnr1, Xnr2, Xnr4 and Derriere) is shown. Edd,
Hnf1ß, Foxa1 and Foxa2 are direct Sox17 target
genes. Siamois, Hex and Cerberus are known ß-catenin
target genes. Xnr1, Xnr2, Xnr4, Derreire and Mixer are
endodermal genes that are not Sox17 targets. Plakoglobin
(Plako) and Ef1
are control genes that are neither
Sox17 nor ß-catenin targets. (B) A proportion of each sample from the
same experiment was assayed by immunoblotting with either anti-ß-catenin
or anti-tubulin antibodies. Injected N-ß-catenin protein has a
higher molecular weight than endogenous ß-catenin because of the presence
of an epitope tag.
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