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Fig. 7. The N-terminal extracellular domain of Xfz3 is unable to inhibit Xfz3
dimerization and activation of the Wnt/ß-catenin pathway in response to
overexpression of Xfz3 in Xenopus embryo. Embryos were injected with
mRNA encoding Xfz7-myc, Xfz3-HA or a combination of Xfz3-HA and extra3-myc
mRNA. (A) RT-PCR analysis of the Wnt/ß-catenin target gene
siamois in animal caps cultured to the early gastrula stage.
Siamois expression in whole embryo is used as a positive control and
in noninjected caps as a negative control. ODC is a loading control.
(B) Immunoprecipitation and western blot analysis of protein extracts from
injected embryos. Expression of Xfz3-HA is revealed by immunoprecipitation
using the anti-HA antibody followed by western blot analysis with the same
antibody. The dimeric form of Xfz3-HA (arrow) is detected after injection of
Xfz3-HA mRNA alone (lane 2) or in combination with extra3-myc mRNA (lane3).
Expression of the extra3-myc protein in co-injected embryos is checked by
immunoprecipitation and western blot with the anti-myc antibody (lane 5,
arrowhead).