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Fig. 7. Sox17ß requires ß-catenin to robustly activate target gene
transcription in animal caps. (A) Embryos were injected at the two-cell stage
with the indicated combinations of: Sox17ß mRNA (200 pg), RNA encoding a
N-terminal deleted form of stabilized ß-catenin (
N-ß-catenin,
100 pg) (Yost et al., 1996),
or an antisense ß-catenin morpholino oligos (ßcat-MO; 20 ng). At
blastula stage, animal cap tissue was explanted and cultured for 3-4 hours
until gastrula stage, when it was assayed by real-time RT-PCR for the
expression of Sox17 target genes. The histograms show the relative expression
levels normalized to the loading control, ODC. Plakoglobin
(Plako) is control gene that is neither a Sox17 nor ß-catenin
target. (B) A proportion of each sample from the same experiment was assayed
by immunoblotting with anti-ß-catenin, anti-Sox17ß 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. Tubulin is a loading control.
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