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Fig. 4
Role of SoxB binding sites in the regulation of cyp26a1 expression in the anterior neural plate (ANP).(A) Representation of the cANE region in the zebrafish genome; 3 predicted SoxB binding sites are indicated; the green channel represents the intensity of the anti-Sox2 ChipSeq signal in this region according to [34]. (B) A logo representing the composite consensus binding site for SoxB/Oct factors [33] is aligned with the predicted Sox binding site (Sox_BS1) overlapping Motif1 (cANE 45-59). The mutSox TT->CC mutation destroying the Sox-binding half-site is indicated. (C-F) egfp in situ hybridization representing enhancer activity of cANE ΔSoxBS2-3, where both Sox_BS2 and Sox_BS3 have been deleted (C), compared with intact cANE (D), at the 90% epiboly stage, and enhancer activity of Motif1 mutation mutSox (F) in 1-222 context compared with wild type 1-222 (E), at the 75% epiboly stage. Dorsal views; anterior is to the left. (G) RT-PCR relative quantification of total egfp expression stable transgenic embryos for constructs cANE (1-310), 81-310, 1-222 and 1-222 mutSox, as well as non transgenic embryos (Ctl); error bars represent SEM; units are arbitrary.