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. Author manuscript; available in PMC: 2009 Jun 18.
Published in final edited form as: Cancer Lett. 2008 Mar 17;264(2):265–273. doi: 10.1016/j.canlet.2008.01

Figure 4. ChIP Assay for Caveolin-1 Promoter.

Figure 4

Panel A. Formaldehyde fixed MCF-7 cell chromatin was digested (17.5′) with Enzymatic Shearing Cocktail (Active Motif). Following cross-link reversal and proteinase K treatment, sheared DNA was electrophoresed on 1% agarose and stained with ethidium bromide. The gel contains a 100 bp molecular weight ladder (MWM), and DNA that yielded 100–200 bp fragments suitable for ChIP assays. Panel B (ChIP Assay). Following the immunoprecipitation chromatin from EtOH (controls) or THF-diol treated MCF-7 cells, the DNA was subjected to PCR analysis (36 cycles) with primers for the CAV1 promoter. PCR products from the Input DNA (1:10 dilution) are shown as additional controls. Results from 3 PCR reactions (as in panel B) for EtOH controls or THF-diol treated cells were quantified with UN-SCAN-IT (Silk Scientific Software), normalized to Input DNA and analyzed with Instat (panel C).