Ets-2-binding sites contribute to promoter activation of the UCA1.

<p>(A) Luciferase assay. The Ets-2 binding sites (GGGAAG) were replaced with <i>Hind</i>III sites (AAGCTT) by site-directed mutagenesis (upper panel). Relative luciferase activities of the wild-type (wt) and the mutant vectors (mutant) were measured by luciferase assay. Values represent means ± SD of at least three independent experiments. *<i>P</i><0.05. (B) <i>In vitro</i> detection of Ets-2 transcription factor binding to the promoter region of the UCA1 gene by EMSA assay. One predominant band was observed when biotin-labeled Ets-2 probe was incubated with the nuclear extract (lane 2). Ets-2 binding could not be inhibited by the mutant probe (lane 3) while it was competed with the unlabeled cold probe (lane 4). (C) <i>In vivo</i> detection of Ets-2 transcription factor binding to the promoter region of the UCA1 gene promoter by ChIP assay. Input chromatin (Input), which represented portions of sonicated chromatin before immunoprecipitation, was used as a positive control. IgG antibody was used as a negtive control of ChIP assay.