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

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(A). Structure of ABCB4 CpG island promoter on chromosome 7q21.12 and the ACBC4 protein. Arrow marks transcriptional (+1) start site and boxes mark exons of ABCB4. The ABCB4 protein consists of two ABC transporter transmembrane regions (ABC_membrane) and two ATP-binding cassette domains (ATB_BCD). (B). ABCB4 levels in normal tissues. Expression of ABCB4 was analyzed in RNA isolated from normal lung, breast, liver and kidney samples by real-time RTPCR and normalized to GAPDH mRNA levels. (C). Promoter assay. HEK293 cells were transfected with 1 μg of pRL-null, pRL-ABCB4 and in vitro methylated (ivm) pRL-ABCB4 promoter construct and 0.35 μg of pGL3. Cells were isolated 24 h after transfection and studied using a dual-luciferase reporter assay.