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. 2016 Jul 16;8(32):52104–52117. doi: 10.18632/oncotarget.10631

Figure 4. Methylation level of miR-375 promoter under DAC treatment.

Figure 4

(A) Schematic illustration of the methylation-specific qPCR (MSP) assay performed to quantify the methylation level in the miR-375 promoter. The graphic shows six CpG dinucleotides located in the miR-375 promoter, which were covered by the primers used for MSP. These primers were designed to amplify bisulfite converted and methylated template DNA. (B) Comparison of the methylation level in the miR-375 promoter region between HPV-transformed cell lines treated with 0.5 μM DAC and DMSO using MSP analysis. The bisulfite and methylation-specific primers were designed to cover CpG dinucleotides located in close proximity to the start of the miR-375 gene (as shown in A). As a reference gene bisulfite converted and unmethylated β-actin sequences were amplified indicating successful bisulfite conversion and sufficient DNA quality. The results are presented as mean methylation levels from at least three independent treatments. The error bars indicate the according standard deviation and Student`s t-test was used to calculate p values. *p < 0.05 and **p < 0.01.