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

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(A). Genomic organization of Abcb4 CpG island on chromosome 5qA1. Transcriptional start, CpG sites and COBRA primers are depicted. The PCR product (148 bp) with the TaqI site (pos. 42) is indicated. (B). Methylation analysis of Abcb4 by COBRA. Bisulfite converted DNA was amplified and PCR products were digested with TaqI (+) or mock-treated (−). Methylation status of Abcb4 is indicated (m = methylated, u = unmethylated). (C). Expression analysis of murine Abcb4 in heart, liver, lung and kidney and in murine cancer cell lines LLC1 and Ltk-. Expression of Abcb4 (200 bp) and Gapdh (156 bp) was analyzed by RT-PCR on a 2% gel.