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. Author manuscript; available in PMC: 2007 Apr 1.
Published in final edited form as: Gastroenterology. 2006 Apr;130(4):1283–1300. doi: 10.1053/j.gastro.2006.01.010

Fig. 5. Chromatin Immunoprecipitation assay demonstrates that HNF6 and Foxm1 proteins bind to the endogenous TGFα and Cyclin D1 promoter regions.

Fig. 5

The mouse TGFα promoter region contains distinct DNA binding sequences for the HNF6 protein (−1608 to −1591 bp) and the FoxM1 protein (−1561 to −1531 bp), whereas these transcription factors bind to the same DNA sequence in the mouse Cyclin D1 promoter region (−1064 to −1049 bp). Mouse hepatoma Hepa1-6 cells were mock infected or infected with AdHNF6 or adenovirus expressing Green Fluorescent Protein (AdGFP). For the Cyclin D1 promoter ChIP assays (Panel D), we also infected Hepa1-6 cells with AdHNF6 or AdM1b (adenovirus expressing human FoxM1b) separately or together. At 24 hours after infection the chromatin was cross-linked, sonicated to DNA fragments of 500 to 1000 nucleotides in length, and then processed for ChIP assays as described in the Materials and Methods. The cross-linked and sonicated mouse chromatin was immunoprecipitated (IP) with antibodies specific to either HNF6 (+, A and B) or FoxM1 (+, C and D) or with rabbit antisera (−) and the amount of promoter DNA associated with the IP chromatin was quantitated by Real-Time PCR with primers specific to either the −1647 to −1495 bp mouse TGFα promoter region (A and C) or the −890 to −733 bp mouse Cyclin D1 promoter region (B and D). The asterisks indicate statistically significant changes: *P ≤ 0.05, **P ≤ 0.01 and P ≤ 0.001.