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. Author manuscript; available in PMC: 2011 Jan 1.
Published in final edited form as: Breast Cancer Res Treat. 2009 Feb 11;119(1):63–70. doi: 10.1007/s10549-009-0330-4

Figure 3. Ets1 binds to the αB-crystallin promoter in vitro and in vivo.

Figure 3

(A) EMSA analysis of Ets1 binding to the putative EBS in the human αB-crystallin promoter in vitro. The human αB-crystallin promoter region containing the putative WT EBS or a mutant (Mut) EBS was duplexed with the corresponding antisense oligonucleotide and used as probes. Twenty fmol of biotinylated WT or mutant probe was added to nuclear extract in the absence or presence of 4 pmol of unbiotinylated WT probe. For antibody abrogation experiments, the binding reaction was preincubated with an Ets1 antibody or control IgG before adding the WT probe. Protein-DNA complexes were resolved by native-PAGE, transferred to a membrane, and detected by chemiluminescence. (B) ChIP analysis of endogenous Ets1 binding to the human αB-crystallin promoter in vivo. Input DNA-protein complexes or DNA-protein complexes immunoprecipitated with water, IgG or Ets1 antibody were PCR amplified using primers flanking the αB-crystallin promoter EBS.