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. Author manuscript; available in PMC: 2013 Feb 1.
Published in final edited form as: Arch Biochem Biophys. 2011 Dec 16;518(1):79–88. doi: 10.1016/j.abb.2011.12.006

Figure 6. ChIP analysis indicates acetylation of histones on the NF-κB2 promoter.

Figure 6

ChIP analysis was performed on H1299 cells stably transfected with vector (HC5) or mutant p53-R273H (R273H) to test whether histones are preferentially acetylated on the NF-κB2 promoter, and if different transcription factors and mutant p53 are nucleated on the promoter. As described in Materials and methods, cells were crosslinked with formaldehyde, harvested and DNA sonicated to generate 500 bp-2 Kb fragments. Designated proteins were immunoprecipitated using respective antibodies. Cross-linking was reversed and the DNA isolated. QPCR was performed using gene specific primers. Normalized values represent QPCR values normalized to the GAPDH gene whose expression is not significantly affected by mutant p53. The promoter specific primers are located within 1000 bp of the transcriptional start site. A. ChIP performed with antibodies directed against acetylated histone H3 (acetylated at K9 and K14) and H4 (acetylated at K16). B. ChIP performed with antibodies directed against CREB, NF-κB p65, and STAT. C. ChIP performed with antibodies directed against CBP, cRel and p53 DO1. D. ChIP performed with an antibody against the total p53 protein.