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. 2010 Apr 1;21(7):1166–1177. doi: 10.1091/mbc.E09-08-0689

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© 2010 by The American Society for Cell Biology

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Figure 6. — I3C disrupts ERα protein interaction with GATA3 regulatory regions. (A) Genomic sequences of the GATA3 gene enhancer contain a consensus half-ERE site. Primers used to amplify ERE site for chromatin immunoprecipitation are underlined. Sequence and chromosomal location were obtained from the UCSC Genome Browser. (B) ChIP was used to characterize endogenous ERα interactions with the ERE region of the GATA3 enhancer region. Chromatin was isolated from MCF-7 cells treated with or without 200 μM I3C for 24 h. ERα was immunoprecipitated from total cell extracts using Sepharose G bound to anti-ERα antibody, and DNA released from ERα was amplified using the indicated oligonucleotide primers. Control primers directed at downstream site (distance, 1256 base pairs) showed no amplification in immunoprecipitated (IP) samples. Input samples represent total genomic DNA from each treatment (loading control). This result was repeated twice.