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Fig. S5
Ephb4-2 enhancer activity requires SMAD4. a. X-gal expression in comparably sized Smad4EC/EC;Ephb4-2:LacZ and Smad4+/+;Ephb4-2:LacZ transgenic embryos. b. Representative transverse sections taken through the torso at the base of the heart from E10.5 Smad4+/+ and Smad4EC/EC embryos also transgenic for Ephb4- 2:LacZ. Cv indicates cardinal vein, da indicates dorsal aorta (section shown for Smad4EC/EC embryo can be directly compared to Fig. 1b to confirm da identification). c. Table summarizing the reduction of GFP expression pattern in transient transgenic zebrafish after mutations of SMAD binding elements (SBE), and other conserved regions (CONT), comparative to Ephb4-2:GFP wild-type. The numbers after SBE and EBE correlate to the motifs highlighted in Supplementary Fig. 4c, mutated nt in CONT are marked in orange text in Supplementary Fig. 4c. d. Representative 48hpf transient transgenic zebrafish expressing wild type and mutated Ephb4-2 sequences detailed in c. e. Table summarizing the reduction of X-gal expression pattern in transient transgenic mice expressing Ephb4-2mutSBE(2,3/6/7):LacZ, in which two composite SMAD binding motifs have been mutated, compared to Ephb4-2:LacZ wild-type.