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. 2001 Sep 1;21(17):6810–6819. doi: 10.1523/JNEUROSCI.21-17-06810.2001

Fig. 3.

Fig. 3.

The Pitx2 protein binds to sequences in the mouseGad1 promoter. A, The sequence of the mouse Gad1 promoter region containing the Pitx2 binding sites. The first nucleotide of this sequence is 246 nucleotides upstream of the previously defined major transcriptional start site (Szabo et al., 1996). Underlining indicates the nucleotides that are conserved between the mouse and humanGad1 genes. The Pitx binding sites are shown inlarger letters. B, C, The 5′ site (B) and 3′ site (C) bind to Pitx2 protein specifically. Shifted protein-DNA complexes are indicated by arrows. Reactions containing probe only (Probe), 250-fold or 500-fold excess of mutant competitors (M250, M500), or purified GST protein (GST) are indicated. Increasing amounts (50- to 200-fold molar excess) of unlabeled 5′ site were added as a specific competitor as indicated by the wedges. A bicoid binding site from the Drosophila hunchback promoter was used as a positive control (lanes markedhunchback). The binding of GST-Pitx2 protein to the hunchback promoter site was also competed away by the sites from the mouse Gad1 gene.