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. Author manuscript; available in PMC: 2009 Feb 8.

Published in final edited form as: Brain Res. 2007 Dec 5;1193:1–11. doi: 10.1016/j.brainres.2007.11.055

Egr1 in vitro interacts with the putative Egr1 binding motif on the DBH promoter. (A) ³²P-labeled DBH Egr1 double-stranded oligonucleotide (−243/−210) was incubated with nuclear extract from pCMV Egr1-transfected cells alone (lane 2), with 100 × excess unlabeled double-stranded oligonucleotide (lane 3), or with 4- or 2 μg specific Egr1 antibody (lanes 4 and 5). Lane 1 is the labeled DBH Egr1 oligonucleotide alone. Bands are indicated by * to show the displacement of the bands by Egr1 antibody. (B) ³²P-labeled double-stranded consensus Egr1 oligonucleotide was incubated with nuclear extracts from pCMV Egr1-transfected cells alone (lane 2), with 100 × excess unlabeled double-stranded consensus oligonucleotide (lane 3), with 100 × excess unlabeled double-stranded mutated consensus Egr1 oligonucleotide (lane 4), or with 700 × excess unlabeled double-stranded DBH Egr1 oligonucleotide (−243/−210) (lane 5). Lane 1 is the labeled consensus Egr1 oligonucleotide alone. Bands are indicated by * to show the displacement by consensus Egr1 and DBH Egr1 oligonucleotides (−243/−210), but not by the oligonucleotide with mutated Egr1. Right panel is the amplified image of the representative gel.

Egr1 in vitro interacts with the putative Egr1 binding motif on the DBH promoter. (A) ³²P-labeled DBH Egr1 double-stranded oligonucleotide (−243/−210) was incubated with nuclear extract from pCMV Egr1-transfected cells alone (lane 2), with 100 × excess unlabeled double-stranded oligonucleotide (lane 3), or with 4- or 2 μg specific Egr1 antibody (lanes 4 and 5). Lane 1 is the labeled DBH Egr1 oligonucleotide alone. Bands are indicated by * to show the displacement of the bands by Egr1 antibody. (B) ³²P-labeled double-stranded consensus Egr1 oligonucleotide was incubated with nuclear extracts from pCMV Egr1-transfected cells alone (lane 2), with 100 × excess unlabeled double-stranded consensus oligonucleotide (lane 3), with 100 × excess unlabeled double-stranded mutated consensus Egr1 oligonucleotide (lane 4), or with 700 × excess unlabeled double-stranded DBH Egr1 oligonucleotide (−243/−210) (lane 5). Lane 1 is the labeled consensus Egr1 oligonucleotide alone. Bands are indicated by * to show the displacement by consensus Egr1 and DBH Egr1 oligonucleotides (−243/−210), but not by the oligonucleotide with mutated Egr1. Right panel is the amplified image of the representative gel.