Figure 4.

Methylation analysis in single glands with and without IM. A: Genomic structure of the putative promoter region of ZIK1. The region was divided into X and Y regions for bisulfite sequencing. Closed box, DNA fragment isolated by MS-RDA; arrowheads, locations of MSP primers. B: Representative photos of single glands with and without IM isolated by single gland isolation technique. C: Results of bisulfite sequencing of single glands in six cases. Gastric glands were isolated from the anterior and posterior walls of the six cases, and the presence or absence of IM was determined by Alcian blue staining. DNA was extracted from a single gland, and bisulfite sequencing was performed. The vast majority of DNA molecules obtained from a single gland with IM were methylated, whereas those from a single gland without IM were unmethylated. Closed circle, methylated CpG site; open circle, unmethylated CpG site.