| |
Fig. 4. ASPN is required for eye
development. (A-C) Representative images
from the injection of control-MO (A), ASPNMO1
(B) and ASPN-MO1 together with the
coding region of ASPN (ASPNCDR) mRNA (C).
(D) Quantification of the phenotypes. For the
rescue experiment, embryos were injected
with either 20 ng ASPN-MO1 and 1 ng
ASPNCDR, or 20 ng ASPN-MO1 and 3 ng
ASPNCDR, and the phenotypes analysed at
stage 41. (E-X) Expression of marker genes
caused by ASPN-MO1. Either control-MO
(E,G,I,K,M,O,Q,S,U,W) or ASPN-MO
(F,H,J,L,N,P,R,T,V,X) was injected together
with β-Galactosidase mRNA as a tracer (light
blue product) and embryos were analysed at
stage 17 (E,F,I,J,M,N,Q,R,U,V) or stage 22
(G,H,K,L,O,P,S,T,W,X) by in situ hybridisation
with the probes of Rx (E,F), Six3 (G,H), Pax6
(I,J), Six6 (K,L), Otx2 (M-P), En2 (Q-T) and
Krox20 (U-X). Arrowheads in F and J indicate
affected areas. (Y) ASPN is essential for the
induction of EFTFs by Chordin (Chd). Animal
caps of control (i; black bars), Chd-injected
(ii; blue bars) and Chd+ASPN-MO-injected
(iii; red bars) embryos were prepared and the
animal caps were analysed at stage 22 by
qRT-PCR (*P<0.01; Student’s t-test).
Error bars represent s.e.m. |
