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Fig. 7. XMeis3 caudalizing activity functions in the absence of canonical Wnt
signaling. All embryos were injected in the animal hemisphere with RNA or DNA
at the one-cell stage and animal caps were removed at blastula stages 8-9. (A)
XMeis3/AC control recombinant explants (no elongations, n=0/28). (B)
XMeis3/BMP DNR recombinant explants, in which the neuralized albino cells
undergo elongation (83% elongations, n=25/30). (C) XMeis3/BMP DNR
explants co-injected in the albino neuralized explant with
ß-catenin MO. These explants elongate (85% elongation,
n=17/20). (D) Albino explants were also co-injected with the
ß-catenin-promoter-driven luciferase reporter construct 3X
TCF-luc on the albino side as described in (A,B), except that 0.5 ng
GSK-3-encoding RNA was injected instead of ß-catenin MO. At
early neurula stages, 8-15 explants were lysed per group and luciferase
activity assayed. The graph shows relative luciferase activity in each sample,
with the control (A) taken as 100% luciferase activity. (E) One-cell-stage
embryos were injected in the animal hemisphere with 1.0 ng XMeis3
RNA, 0.5 ng GSK-3 RNA or both. 18 animal cap explants were removed
from uninjected and injected groups of blastula embryos (stage 8-9). Explants
from each group were grown to stage 20 and total RNA was isolated. RT-PCR
analysis was performed with the markers Krox20 and HoxB9.
EF1
served as a control to quantify RNA levels in the different
samples. For controls, RT-PCR and –RT-PCR was performed on total RNA
isolated from normal embryos.
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