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¹ Cellular Biochemistry, Animal Resource Sciences/Veterinary Medical Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan
² Departments of Medicine (Hematology) and Molecular Genetics, Albert Einstein College of Medicine, Bronx, NY, USA

Our previous studies using restriction landmark genomic scanning (RLGS) defined tissue- or cell-specific DNA methylation profiles. It remains to be determined whether the DNA sequence compositions in the genomic contexts of the NotI loci tested by RLGS influence their tendency to change with differentiation. We carried out 3834 methylation measurements consisting of 213 NotI loci in the mouse genome in 18 different tissues and cell types, using quantitative real-time PCR based on a VIRTUAL IMAGE RLGS database. Loci were categorized as CpG islands or other, and as unique or repetitive sequences, each category being associated with a variety of methylation categories. Strikingly, the tissue-dependently and differentially methylated regions (T-DMRs) were disproportionately distributed in the non-CpG island loci. These loci were located not only in 5'-upstream regions of genes but also in intronic and non-genic regions. Hierarchical clustering of the methylation profiles could be used to define developmental similarity and cellular phenotypes. The results show that distinctive tissue- and cell type-specific methylation profiles by RLGS occur mostly at NotI sites located at non-CpG island sequences, which delineate developmental similarity of different cell types. The finding indicates the power of NotI methylation profiles in evaluating the relatedness of different cell types.

^aThese authors contributed equally to the work.

^bPresent address: Pfizer Japan Inc., Shinjuku Bunka Quint, 3-22-7 Yoyogi, Tokyo 151-8589, Japan.

^cPresent address: Department of Medicine (Hematology), Albert Einstein College of Medicine, Bronx, NY, 1046 USA.

^* Correspondence: E-mail: ashiota{at}mail.ecc.u-tokyo.ac.jp

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