Fig. 4. (A) Sequence of the 5' flanking region between nucleotide positions –1142 and –963 of Cs-FoxD, which contains three Tcf-binding motifs [TBE1-3: CTTTG(A/T)(A/T)]. (B) Relative levels of lacZ expression when various deletion constructs were injected. Deletion constructs are shown on the left and relative ratios of lacZ expression on the right. The relative expression level was calculated as described in the Materials and Methods. (C) Relative levels of lacZ expression in embryos injected with ß-catenin mRNA, cadherin mRNA or various mutation constructs derived from p(–1138)Cs-FoxD/lacZ. Mutation N is shown in A. (D) Expression of lacZ when control p(–1138)Cs-FoxD/lacZ was injected. lacZ was detected in progeny cells of A5.1, A5.2 and B5.2 (endoderm, notochord and nerve cord cells, respectively) presumably due to the stability of ß-gal protein. (E) Expression of lacZ was not detected when p(–1138/del TBE1/2/3)Cs-FoxD/lacZ was injected.