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Fig. 2. Direct targets of Sox17ß. (A) A schematic of the GR:Sox17ß fusion
protein consisting of the hormone-binding domain of the human glucocorticoid
receptor fused to Sox17ß. (B) Isolated blastula animal cap explants
either injected with mRNA encoding GR:Sox17ß (150 pg) or uninjected were
each cultured in three conditions: 1xMBS, 1xMBS +
10–6 dexamethasone (DEX) or in 1xMBS with
10–6 dexamethasone + 10 µg/ml cycloheximide (CHX).
Dexamethasone activates the GR:Sox17ß fusion protein and cycloheximide
blocks translation. Control animal caps were treated with 5 ng/ml human
activin A either with or without 10 µg/ml cycloheximide. At gastrula (stage
11) the explants were assayed by RT-PCR. GR:Sox17ß induced
Hnf1ß, Foxa1, Foxa2 and Sox17
transcription when translation was blocked and are therefore direct Sox17
targets. A repeat of this experiment assayed by real-time RT-PCR is presented
in Figs S1-S3 at
http://dev.biologists.org/supplemental.
(C) Whole-mount in situ hybridization to bisected gastrula (stage 11) embryos
with the indicated probes confirms that Sox17 target genes are expressed in
the endoderm. Dorsal/anterior towards the left.
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