XB-IMG-153860
Xenbase Image ID: 153860
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Fig. 5. Cooperation of ASPN and IGF is essential for eye
development. (A) ASPN activates ERK and AKT.
Conditioned media taken from control GFP (lane 1), ASPN
(lane 2) or IGF2 (lane 3) expressing cells were applied to HEK
293 cells for 20 min.Western blotting analysis was performed
with antibodies for phosphorylated ERK (p-ERK), ERK,
phosphorylated AKT (p-AKT) and AKT. (B) ASPN physically
interacts with IGF1R. HEK 293 cells were transfected with
expression vectors carrying IGF1R (lanes 1,2) and ASPN
(lane 2) and co-immunoprecipitation analysis was performed
with the IGF1R antibody and detected with the myc antibody.
IB, immunoblotting; IP, immunoprecipitation. (C-I) Embryonic
eye formation requires both ASPN and IGF signals.
(C-G) Embryos were injected with 3 ng β-Galactosidase
mRNA (control: C), 1 ng ASPN mRNA (D), 1 ng ASPN+3 ng
dnIGFR mRNAs (E), 1 ng IGF2 mRNA+20 ng control-MO
(F) or 1 ng IGF2 mRNA+10 ng ASPN-MO (G) into the dorsal
blastomere at the 4-cell stage and phenotypes were evaluated
at stage 42. Affected areas are indicated with yellow
arrowheads. (H,I) The same combination of mRNAs and
morpholinos were injected. Animal caps were prepared and
analysed at stage 22 for Pax6 and Rx2a expression with
qRT-PCR (*P<0.01; Student’s t-test). Error bars represent
s.e.m. Image published in: Luehders K et al. (2015) Copyright © 2015. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View |