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Fig. 3. In vitro analysis of Cv-2 activity. (A) Western blot analysis and
co-immunoprecipitation reveals the direct interaction of V5-tagged Cv-2
protein with recombinant human BMP4 protein. V5-tagged chordin is used as a
positive control for BMP binding. (B-E) Effect of Cv-2 injection on axis
formation in Xenopus at stage 14 (B) and stage 27 (C-E). Ventral
injections of Cv-2 RNA induce a partial secondary axis (arrows; B,C,E).
Transverse section through the trunk of a stage 27 embryo injected ventrally
with Cv-2 confirms the presence of an ectopic secondary neural epithelium
(arrow, D). (F,G) RT-PCR analyses of Xenopus animal cap explants. (F)
Xenopus Noggin mRNA induces expression of the general neural marker
NCAM and anterior neural marker BF1. Increasing amounts of Cv-2 mRNA
do not induce expression of either neural marker. (G) Co-injection of 2 ng
Cv-2 RNA plus 0.1 ng noggin RNA increases the induction of BF-1 when compared
with injection of 0.1 ng noggin RNA alone. The mesodermal marker muscle actin
is used to confirm the absence of mesoderm in the explants and elongation
factor-1 alpha (EF-1
) is used to assess the relative amounts
of recovered mRNA.
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