(A–E) Human ski neuralizes Xenopus
ectoderm. Explants were cultured until stage 20 for analysis of
NCAM expression. (A) Embryos express NCAM
only in the central nervous system. Anterior is to the left and dorsal
is to the top. (B) Uninjected explants.
(C) Explants expressing 500 pg of BF-2 mRNA.
(D) Explants expressing 500 pg of human ski
mRNA. (E) RT-PCR analysis. Lanes 1 and 2, whole
embryo control (stage 20) followed by the RT minus negative control;
lane 3, uninjected explants; lanes 4–6, explants injected with
increasing amounts of ski mRNA. The elongation factor 1
alpha (EF1α) primers were used to determine RNA recovery. Primers for
muscle actin were used to assess mesoderm development. W,
whole embryo; RT−, no RT. (F–I) Ski inhibits
Xvent2 expression in ectoderm. Ectodermal explants were
cultured until the midgastrula stage (stage 10.5). (F)
In the embryo, Xvent2 is expressed in the animal cap and
the marginal zone (dorsal lip is indicated by the arrow).
(G) Uninjected explants. (H) Explants
expressing 500 pg of BF-2 mRNA. (I) Explants
expressing 500 pg of ski mRNA. (J–M) Ski
inhibits Xvent2 expression in the embryo.
(J) Control embryos (ventral view) showing
Xvent2 expression in the marginal zone (stage 10.5).
(K) Embryos injected with 500 pg of ski mRNA;
arrowheads point to where Xvent2 expression is
inhibited. (L) Control embryos probed for noggin
expression. (M) Embryos injected with 500 pg of
ski mRNA probed for noggin expression.
(N–Q) Ski does not inhibit BMP4 expression
in the ectoderm. Explants were cultured until the midgastrula stage
(stage 10.5). (N) In the embryo, BMP4 is
expressed strongly in the ectoderm and the marginal zone (dorsal lip is
indicated by the arrow). (O) Uninjected ectodermal
explants. (P) Explants expressing 500 pg of
BF-2. (Q) Explants expressing 500 pg of
ski.