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Fig. 1. (A) Overexpression of FRL-1 induced the expression of neural
markers without mesodermal markers in animal caps. 1 ng of FRL-1 or
100 pg of chordin (chd) were injected into animal caps,
which were dissected at stage 9, together with an uninjected control, and
harvested at stage 13 with the whole embryos for RT-PCR analysis.
FRL-1 induced the expression of zic3, soxD, Xngnr-1 without
induction of Xbra and chordin, and inhibited Xmsx-1
and Xvent-1. `ND' indicated no data. (B) FRL-1 inhibits BMP
signaling in animal caps. Animal caps injected with 1 ng of BMP-4
with or without 1 ng of FRL-1, were dissected at stage 9 and
harvested at stage 10.5 for RT-PCR. The induction of Xvent-1, Xmsx-1
and Xbra induced by BMP-4 was inhibited by co-injection of
FRL-1, and the inhibition of zic3 and soxD by
BMP-4 was rescued by co-injection of FRL-1. (C)
FRL-1 induced the expression of neural markers, N-CAM and
otx2 in a concentration-dependent manner without induction of
mesodermal markers, muscle specific-actin (ms-actin) and
collagen type II (col II). 1 ng or 5 ng of
FRL-1-injected animal caps were dissected at stage 9 with the
uninjected control and harvested at stage 26 for RT-PCR.
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