Spacing between EBSs constrains specific EIN3 binding.

<p>(A) Reported natural promoters of EIN3 targets. EBSs are shown in red. The spacing sequences between two EBSs are underlined and marked with the spacing lengths. All listed EIN3 targets have been proved by EMSA. (B) DNA-binding of EIN3 to probes with varying length of the spacing between EBSs. The sequences of the probes are listed in the lower panel. NC represents “negative control”, which is the free probe of <i>2EBS-S10</i> without the addition of any protein. (C) Relative binding data for 0.2 μM EIN3 82–352 to the probes in (B). The data were normalized to the fraction bound of <i>2EBS-S10</i>; error bars represented the standard deviation from three independent experiments. (D) Quantified binding data and (E) dissociation constants for probe <i>2EBS-S5</i>, <i>2EBS-S8</i>, and <i>2EBS-S10</i>. <i>K</i>_d values were obtained from three independent experiments.