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. 2015 Jul 23;65(10):1619–1631. doi: 10.1136/gutjnl-2015-309276

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MDGA2 is inactivated by promoter methylation in gastric cancer cell lines. (A) MDGA2 was silenced or downregulated in 10 out of 11 gastric cancer cell lines but readily expression in normal gastric tissue. The methylation status of MDGA2 was determined by methylation-specific PCR (MSP). M, methylated; U, unmethylated. (B) A typical CpG island is present at the promoter region of MDGA2. Each vertical bar represents a single CpG site. The transcription start site (TSS) is indicated by a curved arrow. A region for bisulfite genomic sequencing (BGS) and combined bisulfite restriction analysis (COBRA) is denoted. BGS analysis confirmed high levels of promoter methylation in MDGA2-silenced cells and no/low methylation in MDGA2-expressing samples. (C) MDGA2 mRNA expression was restored after treatment with the demethylation reagent 5-Aza. Decreased methylation was revealed by BGS after 5-Aza treatment.