Changes in DNA methylation of the miRNA locus occurring in an EMT in-vitro model. (A) Phase contrast picture showing differences in cell morphology observed in epithelial HMLE breast cancer cells and in HMLE cells that underwent EMT by constitutively expressing Twist. Scale bar, 10 μm. (B) Methylation status analysis of the region surrounding miR-200c/141 in same cell populations shown in (A). (C) Expression analysis of miR-200c and miR-141 in same cell populations shown in (A). Data are mean of triplicates and are calculated relative to miR-16 expression, which did not exhibit significant differences in Ct values between samples.