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Fig. 3. Functional analysis of FoxD3 fusion proteins. (A) Schematic
of the structure of FoxD3 and the FoxD3 fusion proteins. FoxD3 contains a
conserved winged helix (WH) DNA-binding domain (residues 92-192). The
repressor fusion protein (Eng-FoxD3) contains the repressor domain of
Drosophila Engrailed (residues 1-298) fused to the FoxD3 WH domain.
The activator fusion protein (VP16-FoxD3) contains the activation domain of
HSV VP16 (residues 410-490). Embryos were injected with FoxD3 (100 pg),
Eng-FoxD3 (100 pg) or VP16-FoxD3 (250 pg) and animal explants were analyzed at
the tailbud stage (stage 25) for morphogenesis (B-E) and by RT-PCR for
the expression of Muscle Actin (M. Actin) and Collagen Type
II (Col II) (F). Like FoxD3, Eng-FoxD3 induced convergent
extension movements and mesodermal gene expression, whereas VP16-FoxD3 did
not. (G) Co-expression of VP16-FoxD3 and FoxD3 blocked induction of
mesodermal genes by FoxD3. PCR controls are as described in
Fig. 2.
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