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Fig. 5 | Journal of Experimental & Clinical Cancer Research

Fig. 5

From: DNA methylation of claudin-6 promotes breast cancer cell migration and invasion by recruiting MeCP2 and deacetylating H3Ac and H4Ac

Fig. 5

5-aza-dC may alter the chromatin structure of CLDN6 for transcription in MCF-7 cells. a MCF-7 cells were treated with micrococcal S7 nuclease to produce mononucleosomal genomic DNA fragments (100 ∼ 200 bp). b Primer pairs were designed between -400- + 400 bp of CLDN6 proximal promoter. Micrococcal S7 nuclease preferentially digested DNA that was not organized in nucleosomes. Hence, DNA organized in nucleosomes would be less prone to nuclease digestion and amplified to the qPCR threshold at a lower Ct compared to the less organized DNA. c Purified mononucleosomal DNA was amplified using primer pairs (depicted in (b)) and the resulting product was quantified using SYBR green dye in a quantitative PCR. The difference in Ct between 5-aza-dC treating MCF-7 and DMSO treating MCF-7 cells (as control) for each primer pair was plotted. The positive numbers suggested that in 5-aza-dC treating MCF-7 cells this part of the DNA was less organized into nucleosomes and more prone to nuclease digestion compared to control cells. CLDN6, Claudin-6; 5-aza-dC, 5-Aza-2’-deoxycytidine

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