Fig. 4

miR-200c expression and locus methylation of breast cancer tissues. a miR-200c expression levels of 3 FFPE breast cancers (BrC1, BrC2 and BrC3) and one FFPE normal tissue (BrN) were determined by qRT-PCR and reported as the LOG₂ of the miR-200c levels relative to the RNU48 normalizer control. Data represent the means of three independent experiments performed in triplicate and bars indicate standard deviation. b miR-200c/miR-141 locus PCR of bisulfite treated DNA obtained from the FFPE breast normal tissue (BrN) and cancers (BrC1, BrC2, BrC3). Lane M, 100 bp size marker. NTC, no template control. c Methylation percentages of each CpG in the miR-200c/miR-141 locus of BrC1, BrC2, BrC3 and BrN. Data were obtained with NBSP. d Representative sequencing chromatograms of 6 CpG (highlighted by gray arrows; left panels) and of the reverse sequence of Tail2 of miR-200c/miR-141 amplicon (right panels; with C and T used to calculate the NF indicated by black and white arrows, respectively) for the three breast cancer samples