Kaiso represses expression of the minimal <i>cyclin D1</i> promoter-reporter in a KBS and methyl-CpG dependent manner.

<p>(<b>A</b>) Reporter plasmid methylation was confirmed by digesting the plasmid DNA with the CpG-methylation specific restriction enzyme <i>HpaII</i>. (<b>B</b>) Kaiso overexpression caused a dose-dependent decrease in luciferase gene expression from the minimal <i>cyclin D1</i> promoter reporter possessing active KBS but devoid of methyl-CpG sites (blue bars). Similarly, a dose-dependent decrease was observed when the KBS and CpG sites were both active (red bars). (<b>C</b>) Kaiso overexpression caused dose-dependent repression of luciferase activity in the presence of active methyl-CpG sites and mutated KBSs. (<b>D</b>) Ectopic Kaiso expression had little to no effect on the reporter construct when both the KBS and CpG sites were inactivated.