Skip to main content
. Author manuscript; available in PMC: 2010 Apr 15.
Published in final edited form as: Gene. 2009 Jan 22;435(1-2):36–44. doi: 10.1016/j.gene.2009.01.003

Figure 5. The NFκB site in the jagged-1 promoter binds p50 and p65.

Figure 5

A. EC were treated with TNF (10ng/ml) for 4 hours and nuclear extracts prepared and analyzed by EMSA using a labeled probe covering the NFκB site from the jagged-1 promoter. Where indicated a 50-fold molar excess of unlabeled WT or NFκB mutant probe was added to demonstrate specificity. B. EC were untreated or treated with TNF (10ng/ml) for 4 hours and nuclear extracts prepared and analyzed by EMSA using a labeled probe covering the NFκB site from the jagged-1 promoter. Where indicated a 50-fold molar excess of unlabeled WT or NFκB mutant probe was added to demonstrate specificity. Antibodies to p50, p65, c-rel or control were added to extracts for 20 min. prior to the addition of labeled probe. NS – non-specific band. C. ChIP analysis was performed using control or TNF-treated (10ng/ml) EC and antibodies specific for p50 and p65, or an isotype control. After cross-linking and precipitation, PCR was performed using primers specific for the jagged-1, VCAM-1 or β-actin promoters. Input DNA was used as a positive control.

HHS Vulnerability Disclosure