Fig. 4.

ZEB2 increased DNA methylation at RAB25 promoter through DNMTs activity. a Correlation between RAB25 DNA methylation (x-axis) and ZEB2 (blue dots) or RAB25 (red dots) expression (y-axis) from publicly available NCI-60 cell panel datasets. Mean of each parameter was calculated, individual cell type values reported to the mean and log2 transformed. Pearson’s correlation test was used to calculate r and P values. b Global DNA methylation level upon ZEB2 induction (+dox) in MCF7 was measured by ELISA assay. Control values were set to 1, and s.d. is shown. P values were determined using unpaired t tests (***p < 0.001). c RAB25 promoter DNA methylation in MCF7 was measured by methyl-binding domain (MBD) assay after ZEB2 induction (+dox) and 5′-aza-2-deoxycytidine (+dox/5-aza) treatment. Enrichments to input were calculated, control values were set as 1 and s.d. is shown. P values were determined using one-way ANOVA (*p < 0.05). Three independent experiments were performed