Fig. 3. A novel Zic1 site identified within the Math1 enhancer. Gel shift assays with in vitro transcribed and translated Zic1 (lacking the N-terminal 110 amino acids) and probes generated from (A) a 30 bp sequence from the Math1 enhancer containing the newly identified Zic1-binding site (Z-site wt), or (B) the wild-type 374 bp enhancer B (Math1 enh probe) and Math1 enh probes mutated in the Zic1 site (mutations shown in C). Sequence of the oligonucleotide probe and the cold competitor DNAs are shown in C. Control extract is the reticulocyte extract with no added template. Zic/Gli probe is the previously published Zic consensus binding site. (C) Nucleotide requirements for Zic1 binding defined by EMSA with mutant oligonucleotides are shown. The novel Zic1 site defined here (consensus) has little similarity to the published Zic binding site (Gli site).

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