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Figure 4.
Analysis of Embryos Overexpressing xE2F by Whole-Mount In Situ Hybridization
xE2F RNA was injected into animal poles of eight-cell albino (B) or wild-type embryos (C) and developed to tailbud stages. The expression of HoxB9 was detected in both control and injected embryos by whole-mount in situ hybridization. Anterior is to the left. In (A) and (B), the dorsal view of embryos is shown in the bottom. Arrows indicate position of eyes. Arrowheads indicate ectopic expression sites of HoxB9 in the brain of xE2F-injected embryos. Overexpression of xE2F leads to expansion of HoxB9 expression that is normally restricted in the posterior spinal cord. The ectopic expression of the HoxB9 gene in the brain that is adjacent to eyes is clearly seen in (C). (D–G) Embryos injected with xE2F:GR RNA (2.5 ng) were developed to late blastula stages and then cultured in the presence (E and G) or absence (D and F) of 20 μM DEX. The expression of HoxB9 (D and E) and Otx (F and G) was detected at early neurula stages by whole-mount in situ hybridization. Dorsal views of injected embryos are shown with the anterior side at the top. Location of the blastopore is indicated as bp.
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