Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2009 Jan 7;37(4):1086–1094. doi: 10.1093/nar/gkn1056

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2009 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 2. — In vitro binding of CTCF at the +6.8 kb DHS region. (A) The DHS6.8^oligo and DHS6.8mut^oligo probes. The putative CTCF-binding site core is highlighted. Oligonucleotides were designed to form BssHII sticky ends when annealed (shown in italics), facilitating cloning into the AscI site of pNI. Mutated bases in DHS6.8mut^oligo are underlined. (B) IVT CTCF; (C, D), Caco2 nuclear extracts. (B) EMSA using DHS6.8^oligo. Supershift was performed with an anti-CTCF antibody and anti-RAR β was used as isotype control. (C) EMSA with DHS6.8^oligo. Competition reactions were performed with 100× excess of unlabelled DHS6.8^oligo (self), DHS6.8mut^oligo (self mut), FII (known CTCF-binding site from chicken β-globin locus) and mutant (FIImut). (D) EMSA with DHS6.8^oligo. Supershift reactions were performed as in (B). (E) EMSA with DHS6.8mut^oligo. Complex marked i represents CTCF in complex with DHS6.8^oligo and ii the antibody-supershifted CTCF complex. Undefined interactions formed between 6.8mut^oligo and Caco2 nuclear extract are marked by *.