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Fig. 1. In Xenopus, Krm2 is regulated by Wnt signaling and
expressed in the NC region. (A-D) Whole-mount in situ hybridizations.
(A) Comparison of Xkrm2, XWnt8 and XWnt3a expression
patterns in gastrula and early neurula stage embryos. Top: gastrula stage.
Vegetal view, dorsal is up. Bottom: neurula stage. Dorsal view, anterior is
up. (B) Comparison of Xkrm2 and slug expression
patterns at the indicated stages. Dorsal view, anterior is up. Lowermost
panel: view of frontally cut stage 16 embryos, dorsal is up. Brackets indicate
overlapping expression domains of krm2 and slug.
(C-E) Effect of Wnt pathway perturbations on krm2 expression.
(C) Embryos at the 32-cell stage were injected equatorially in two opposite
blastomeres with 1 ng PPL or dnWnt8 and 250 pg lacZ
mRNA and analyzed at gastrula stage. Arrowheads indicate ß-gal lineage
tracer staining (blue). Vegetal view, dorsal is up. (D) Embryos at the
four-cell stage were injected animally with 200 pg pCS-PPL or
pCSKA-Wnt8 DNA or 100 pg pCS-Wnt3a DNA in one blastomere and
analyzed at gastrula stage. Lateral view, dorsal to the right. (E) Statistical
overview of experiments shown in C and D. (F) Embryos at the four- to
eight-cell stage were injected animally with 100 pg XWnt8 or
Wnt3a mRNA, or 1 or 2 ng BMP4 mRNA. Animal caps were cut at
stage 8-9, cultured until stage 20 equivalent and were analyzed by RT-PCR for
expression of the indicated genes. (G) Embryos at the 32-cell stage
were treated with 120 mM LiCl for 50 minutes, cultured until stage 11.5 and
analyzed by RT-PCR. Histone H4 was used for normalization. -RT, minus reverse
transcriptase control.
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