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Fig. 7. Hex suppresses the activity of Nodal-related TGFß
proteins. (A) Axis duplication phenotypes induced by Hex and
Xnr1. Embryos were injected into a single ventral-blastomere at the four-cell
stage with 500 pg Hex, 250 pg Xnr1 or both and in situ hybridisation carried
out at stage 35 using BF1. (B) Phenotypes induced by Hex and
Xnr1 in ventral marginal zone explants. Embryos were injected with the
indicated RNA at the two-cell stage, explants dissected at stage 10.5 and in
situ hybridisation for BF1 carried out when staging control embryos
reached stage 35. Hex was injected at 500 pg. Arrowheads indicate patches of
BF1 expression. (C) In situ hybridisation for the expression
of Cerberus at stage 10.5. Embryos were injected at the two-cell
stage in both blastomeres with 500 pg Hex and/or 250 pg Xnr1 mRNA. (D)
In situ hybridisation of Hex-depleted embryos for the mesendodermal markers
Cerberus, Goosecoid and Chordin. Embryos were injected as in
Fig. 1F. (E) Hex
suppresses the induction of mesoderm in animal cap explants. Embryos were
injected as in B with 500 pg Hex. Animal caps were dissected at blastula stage
and cultured in 8 U/ml Activin protein until control embryos reached stage 18.
(F) Molecular marker analysis in Hex-injected and Activin-treated
animal cap explants. Animal caps were isolated from embryos injected as in B,
cultured to stage 10.5 in 12 U/ml Activin, and RNA extracted for real-time
RT-PCR analysis for Goosecoid, Chordin, Xbra and Mixer.
Values are normalised to the expression level of Odc. Experiments
were carried out in triplicate.
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