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Fig. 1. Dynamic expression of Xenopus Sebox during gastrulation. A: The expression of sebox and other Mix/Bix genes during early Xenopus
embryonic development, assessed by RT-PCR. The marker odc served as a loading control. B–G: Whole-mount in situ hybridization (WISH) for
sebox at different developmental stages. B–F: Embryos were viewed from the vegetal side, with the dorsal side upward. E0: Sectioned view of a
st11 embryo at the level indicated by the red dashed line in E (d, dorsal; v, ventral). (G) Dorsal view at st14. |
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Fig. 3. Modulation of sebox expression by zygotic Wnt/b-catenin signaling. A: WISH for sebox expression in stage 11 embryos injected with or
without 40 pg Wnt8 plasmid at the two-cell stage. B: WISH for sebox, bix4 and mixer expression in embryos with the indicated treatments.
Embryos were injected with 30 ng b-catenin (b-cat) Morpholino (MO) at the two-cell stage and cultured with or without 100 mM NaCl or LiCl at
stage 8.5 for 30 min. They were then cultured in fresh 0.1 MMR to stage 11, followed by WISH analysis. ND: not determined. C: WISH for sebox,
bix4 and mixer expression in stage 11 embryos injected with 20 ng Wnt8 MO at the two-cell stage. Note that mixer probes were applied to both
intact embryos and half embryos that were bisected along the dorsal–ventral body axis (the bottom row). D: qRT-PCR analysis of the expression
of sebox and mixer in animal caps in response to Activin treatment. Animal caps from 30 ng cMO or b-cat MO-injected embryos were dissected
at st8.5, cultured with Activin (2 ng/ml) for 4 hr and then subjected to RT-PCR analysis. The relative expression of the indicated genes was normalized
to the level of odc. The p values were calculated by a Student’s t-test. n.s. (no significance). E: qRT-PCR analysis of the expression of sebox,
bra, and gsc in animal caps with the indicated treatments. Animal caps were dissected at stage 8.5 from wild-type (ctrl) or b-catenin mRNA (150
pg)-injected embryos and cultured with or without a low dose of Activin (0.1 ng/ml) protein for 4 hr. They were then subjected to qRT-PCR analysis.
The gene expression levels in whole embryos were set to 100%. The relative expression levels of genes were normalized to the levels of odc. |
