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Fig. 7. Activation of PI3K partially rescues eye development in kermit
2/XGIPC-depleted embryos. (A) p110*, a constitutively
active subunit of PI3 kinase, induces AKT phosphorylation/activation in stage
10 animal cap explants. mRNA encoding p110* was injected into
fertilized eggs and animal caps were explanted at the gastrula stage and
analyzed by western blot with antibodies recognizing AKT phosphorylated at
serine-473 (upper panel) or general anti-AKT antibodies (lower panel).
(B) p110* partially rescues eye development in kermit
2-depleted embryos. Kermit 2 morpholino (40 ng), 3 ng of p110*
mRNA, or both, were injected into two dorsal animal blastomeres of
four-cell/eight-cell embryos. No apparent eyes are present in embryos injected
with the kermit 2 morpholino, while small eyes are present in over 70% of
embryos co-injected with p110* mRNA and morpholino.
p110* injection alone does not affect embryo development.
(C) The percentage of embryos with normal, small or absent/miniscule
eyes is summarized. (D) p110* partially recovers the
expression of Xrx in kermit 2-depleted embryos. One dorsal-animal
blastomere of four-cell/eight-cell embryos was injected with kermit 2
morpholino with or without p110* mRNA (right side, indicated by GFP
lineage tracer) and harvested at stage 20. Whole-mount in situ hybridization
was performed for Xrx. Embryos are viewed from the anterior side with
dorsal towards the top.
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