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Fig. 2. Downstream response to the induction of early embryonic signalling by
Hex. (A) Real-time RT-PCR of Cerberus expression in VMZ
explants analysed at stage 10.5. Embryos were injected with the indicated RNA
and Cerberus expression quantified as in
Fig. 1C. Data are based on
three independent experiments. (B) In situ hybridisation for
Cerberus expression and double in situ for Cerberus (purple)
and nucGFP (light blue) expression at stage 10.5. Embryos were
injected with 500 pg Hex, ß-catenin or both at the two-cell stage into
both blastomeres (indicated with a schematic diagram in the lower right-hand
corner) or into a single-ventral blastomere at the four-cell stage (indicated
by `V') alongside a nucGFP RNA. Arrowheads indicate the site of injection.
(C,D) Real-time RT-PCR analysis of Goosecoid and
Chordin. Embryos were injected as in A and RNA from either animal
caps (C) or VMZ (D) explants extracted and analysed at stage 10.5. Values are
normalised to the expression level of Odc and the relative change in
gene expression for the genes analysed was calculated by dividing the values
from injected samples by the values from the uninjected. Data is based on
three independent experiments. (E) In situ hybridisation for
Goosecoid and Chordin expression at stage 10.5. Embryos were
injected with 500 pg Hex, ß-catenin, or both, at the four-cell stage into
a single-ventral blastomere alongside nucGFP. Double staining was performed.
Arrowheads indicate the injected cells. High-magnification views of embryos
co-injected with Hex and ß-catenin are shown for both Goosecoid
(indicated as Gsc) and Chordin (indicated as Chd)
expression. (F) Depletion of Hex by Hex MO. Embryos were injected as in
Fig. 1F and in situ
hybridisation for Cerberus, Goosecoid and Chordin performed
at stage 10.5. Arrowheads indicate the ectopic domain induced by
ß-catenin injection.
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