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. 2014 Nov 4;4:6899. doi: 10.1038/srep06899

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(A). CpG sites at the ABCB4 promoter regions are indicated as vertical lines. The PCR product (218 bp) with two TaqI sites (pos. 70 and 102) and eight CpG sites that were analyzed by pyrosequencing are depicted. (B). Combined bisulfite restriction analysis of ABCB4. Bisulfite-treated DNA from the indicated cancer cell lines and in vitro methylated DNA (ivm) were amplified, digested with TaqI (+) or mock digested (−) and resolved on 2% gels with a 100 bp marker (M); (m = methylated, u = unmethylated). (C). Bisulfite pyrosequencing of ABCB4. Methylation levels of eight CpGs of the bisulfite modified PCR products were analyzed by pyrosequencing.