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Fig. 1. Structure and expression of
ASPN. (A) A phylogenic tree of SLRPs,
which are divided into classes I-V.
(B) Comparison of the amino acid
sequences of ASPN in different species.
The amino-terminal aspartic acid-rich
domain and leucine-rich domains are
circled with red and light blue rectangles,
respectively. Asterisks mark isoleucine,
leucine and valine; black circles mark
cysteine (in the cysteine-rich domain); I-VIII
indicate leucine-rich repeats, as predicted
by a database search using LRR finder
(http://www.lrrfinder.com). (C) Semiquantitative
RT-PCR of ASPN and
Histone4. Whole embryos from various
stages were analysed by RT-PCR. uf,
unfertilised eggs. (D-G) Spatial expression
of ASPN in Xenopus embryos at neurula
stage (st18; anterior view; D) compared
with Pax6 expression at neurula stage
(st18; anterior view; E) and ASPN
expression in early tailbud (st22; lateral
view; F) and tadpole (st35; G) stages was
analysed by in situ hybridisation. The
neural plate border (D) and the
presumptive eye region (F) are indicated by
yellow arrowheads. (H) Expression levels
of ASPN in various types of explants, as
assayed by qRT-PCR. Animal caps
[control (i) or injected with mRNAs of Chd
(ii) and Chd+Wnt8 (iii)] and dorsal marginal
zone (DMZ; iv) and ventral marginal zone
(VMZ; v) were prepared at stage 10.5 and
assayed at stage 18. Error bars represent
s.e.m. |
