| |
Fig. 3. XEpac modulates other hatching gland markers and induces anterior markers. A: Reverse
transcriptase-polymerase chain reaction (RT-PCR) analysis was performed with RNA from XEpac
mRNA (2�4 ng) -injected animal caps for expression of other hatching gland markers. Overexpression
of XEpac induced XAG-1 and XA-1 but reduced XHE, a Xenopus hatching enzyme, and Cx30,
Xenopus connexin30. Additionally, the XOtx-2 anterior neural marker was induced by overexpression
of XEpac. Induction of these markers is irrespective of the induction of Xbra, a pan-mesodermal
marker. Ornithine decarboxylase (ODC) is used as loading control. RT( ), no reverse transcriptase
control; WE, whole embryo. B: XEpac induces XAG-1 and XOtx2 in naive ectoderm and
whole embryo. Two to 4 ng of XEpac were injected into the prospective ectoderm in 2-cell Xenopus
embryos. The injected ectoderm and noninjected control ectoderm were dissected at stage 8.5 and
aged until stage 25. Explants and whole embryos were stained for XAG-1 (a� d) and XOtx2 (e� h) by
in situ hybridization. a, c, e, and g are noninjected control; and b, d, f, and h are injected with XEpac.
Overexpression of XEpac mRNA converted ectoderm into XAG-1- and XOtx2-expressiong cells
and led to the expansion of XAG-1 and XOtx2 expression in the whole embryo. The numbers shown
in a, b, e, f indicate the number of explants exhibiting patches of darkly stained cells per total
number of explants analyzed. |
