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Fig. 4. Generation of a dominant-negative FoxD3 construct. (A) A dominant-negative FoxD3 construct, FoxD3delN. Amino acid residue numbers are indicated above. NLS (grey box), SV40 nuclear localization signal; black box, DNA-binding domain. (B) Animal caps were prepared from embryos injected with control RNA (150 pg), FoxD3 (25 pg; lane 3), FoxD3 (25 pg) + FoxD3delN (50 pg) (lane 4), FoxD3 (100 pg) + FoxD3delN (50 pg) (lane 5), XBF2 (25 pg) + FoxD3delN (50 pg) (lane 6), or XBF2 (25 pg) + FoxD3delN (100 pg) (lane 7) mRNAs. As a specificity control, XBF2delN was also constructed (see Materials and Methods). Animal caps were prepared from embryos injected with XBF2 (25 pg) (lane 8), XBF2 (25 pg) + XBF2delN (50 pg) (lane 9), XBF2 (100 pg) + XBF2delN (50 pg) (lane 10), FoxD3 (25 pg) + XBF2delN (50 pg) (lane 11), or FoxD3 (25 pg) + XBF2delN (100 pg) (lane 12) mRNAs. They were harvested at stage17 equivalent and analyzed by RT-PCR.