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The effect of methylation on the binding of SP1 to CpG#18-20. Labeled oligonucleotides containing unmethylated, singly-methylated, or triple-methylated CpG sites were mixed with nuclear extract derived from 16HBE14o- (A) or CFBE41o- (B) cells and subjected to EMSA. Competition analysis was carried out by adding unlabeled oligonucleotides (lanes 3, 7 and 11) to the reaction solution. Supershift analysis to detect SP1-specific binding was performed using an SP1 antibody (lanes 4, 8 and 12). Arrows indicate the SP1-containing complexes (SP1) and the supershifted SP1 (SS). Bands derived from probes containing 3 methylated CpGs are indicated by (#).
8A, lanes 2, 6, 10).
8A, lanes 4, 8, 12).
8B), suggesting that there is no difference in SP1 binding in
non-CF and CF epithelial cells.
8A, B, lanes 10 and 12, denoted by a sharp).
8A, B, lanes 2, 4, 6 and 8), suggesting that the increased
methylation of CpG#18-20 in non-CF epithelial cells enhances
recruitment of factors that recognize tri-methylated CpGs and
suppress SP1-dependent transcription without affecting the SP1
Furuta, Takashi; Shuto, Tsuyoshi; Shimasaki, Shogo; Ohira, Yuko; Suico, Mary; Gruenert, Dieter C; Kai, HirofumiJournal: BMC Molecular Biology
Issue 1DOI: 10.1186/1471-2199-9-39Published: 2008-12-01Institution(s):
Graduate School of Pharmaceutical Science Global COE "Cell Fate Regulation Research and Education Unit", Kumamoto University, California Pacific Medical Center Research Institute, University of California, San Francisco, University of Vermont
This image is from the article titled "DNA demethylation-dependent enhancement of toll-like receptor-2 gene expression in cystic fibrosis epithelial cells involves SP1-activated transcription"
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