Figure - PMC

Skip to main content

View full-text article in PMC

. Author manuscript; available in PMC: 2013 Jul 16.

Published in final edited form as: Prostate. 2010 Jun 15;70(9):934–951. doi: 10.1002/pros.21128

Fig. 4 — Characterization of the putative Oct-1, C-jun, and NFI binding sites within the –286PB promoter. (A), LNCaP nuclear extracts (10 μg) were incubated with radiolabeled probes corresponding to the wild type or mutated Oct-1 binding sites (−226/−197 bp) in the absence or presence of unlabeled competitor oligonucleotides. The competitors are consensus Oct-1 fragment, wild type Oct-1 binding site, and mutant version of the Oct-1 binding site. Arrows indicate the specific protein-DNA complexes. Complexes A, D, E and F were specifically eliminated or reduced; however, the complex B was enhanced when anti-Oct-1 antibody was added into reaction. Asterisk indicates a supershifted band with anti-Oct-1 antibody. (B), LNCaP nuclear extracts (10 μg) were incubated with radiolabeled probes corresponding to the wild type or mutated C-jun binding sites (−116/−87 bp) in the absence or presence of unlabeled competitor oligonucleotides. The competitors are consensus C-jun fragment, wild type C-jun binding site. Arrows indicate the specific protein-DNA complexes. Complexes A, B, C and D are specifically eliminated or reduced by a consensus C-jun competitor. The protein-DNA complex A was supershifted upon addition of anti-C-jun antibody. Asterisk indicates the supershifted band with anti-C-jun antibody. (C), LNCaP nuclear extracts (10 μg) were incubated with radiolabeled probes corresponding to the wild type or mutated NFI binding sites (−96/−67 bp) in the absence or presence of unlabeled competitor oligonucleotides. The competitors are unlabeled consensus NFI, consensus Oct-1, wt NFI binding site. Two specific protein-DNA complexes are marked as A and B. Both complexes A and B are inhibited upon addition of unlabeled consensus NFI probe. The complex A was supershifted when anti-NFI antibody was included in the reaction. Asterisk indicates a supershifted band with anti-NFI antibody.

Fig. 4 — Characterization of the putative Oct-1, C-jun, and NFI binding sites within the –286PB promoter. (A), LNCaP nuclear extracts (10 μg) were incubated with radiolabeled probes corresponding to the wild type or mutated Oct-1 binding sites (−226/−197 bp) in the absence or presence of unlabeled competitor oligonucleotides. The competitors are consensus Oct-1 fragment, wild type Oct-1 binding site, and mutant version of the Oct-1 binding site. Arrows indicate the specific protein-DNA complexes. Complexes A, D, E and F were specifically eliminated or reduced; however, the complex B was enhanced when anti-Oct-1 antibody was added into reaction. Asterisk indicates a supershifted band with anti-Oct-1 antibody. (B), LNCaP nuclear extracts (10 μg) were incubated with radiolabeled probes corresponding to the wild type or mutated C-jun binding sites (−116/−87 bp) in the absence or presence of unlabeled competitor oligonucleotides. The competitors are consensus C-jun fragment, wild type C-jun binding site. Arrows indicate the specific protein-DNA complexes. Complexes A, B, C and D are specifically eliminated or reduced by a consensus C-jun competitor. The protein-DNA complex A was supershifted upon addition of anti-C-jun antibody. Asterisk indicates the supershifted band with anti-C-jun antibody. (C), LNCaP nuclear extracts (10 μg) were incubated with radiolabeled probes corresponding to the wild type or mutated NFI binding sites (−96/−67 bp) in the absence or presence of unlabeled competitor oligonucleotides. The competitors are unlabeled consensus NFI, consensus Oct-1, wt NFI binding site. Two specific protein-DNA complexes are marked as A and B. Both complexes A and B are inhibited upon addition of unlabeled consensus NFI probe. The complex A was supershifted when anti-NFI antibody was included in the reaction. Asterisk indicates a supershifted band with anti-NFI antibody.