In vivo interaction of SAF-1 transcription factor with VEGF promoter in MDA-MB-231 cells. In vivo interaction of SAF-1 transcription factor with VEGF promoter in MDA-MB-231 cells. HMECs (lanes 1–3 and 7–9) and MDA-MB-231 cells (lanes 4–6 and 10–12) were fixed with formaldehyde and immunoprecipitated with SAF-1 Ab or nonspecific IgG (Nonsp. IgG) following the ChIP assay method as described in Materials and Methods. Immunoprecipitated DNAs were used as templates for PCR amplification of human VEGF promoter by using primers encompassing the SAF-1 DNA–binding element or an unrelated element in the VEGF promoter (20). Amplified products were resolved using a 1.5% agarose gel. Alpana Ray et al. Mol Cancer Res 2011;9:1030-1041 ©2011 by American Association for Cancer Research