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. 2014 Sep 1;35(11):2436–2446. doi: 10.1093/carcin/bgu181

Fig. 2.

Fig. 2.

Analysis of MBD2 expression, promoter methylation, global methylation and transcriptional changes induced by 5-azaCdR, Mbd2 siRNA and combination treatments. (A) DNA was extracted from MCF-7 cells treated with 70nM of siMbd2 and/or 5 µM of 5-azaCdR. Purified DNA was subjected to bisulfite conversion and pyrosequencing of the MBD2 5ʹ region. (B) Quantitative PCR (MCF-7 and ZR-75-1) and western blot analysis (MCF-7) show efficient knockdown of MBD2 in the siMBD2-treated cells and the Combination of 5-azaCdR and siMBD2 treatment. The position of the proteins as determined using molecular weight markers is indicated. (C) Measurement of overall genome DNA methylation at the dinucleotide sequence CpG by nearest neighbor analysis. Percentage methylation is presented as mean ± SEM of triplicate experiments (*P < 0.05, two-tailed Student’s t-test).