Volume 24, Issue 3, Pages (September 2016)

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Volume 24, Issue 3, Pages 502-509 (September 2016) Interindividual Variation in DNA Methylation at a Putative POMC Metastable Epiallele Is Associated with Obesity Peter Kühnen, Daniela Handke, Robert A. Waterland, Branwen J. Hennig, Matt Silver, Anthony J. Fulford, Paula Dominguez-Salas, Sophie E. Moore, Andrew M. Prentice, Joachim Spranger, Anke Hinney, Johannes Hebebrand, Frank L. Heppner, Lena Walzer, Carsten Grötzinger, Jörg Gromoll, Susanna Wiegand, Annette Grüters, Heiko Krude Cell Metabolism Volume 24, Issue 3, Pages 502-509 (September 2016) DOI: 10.1016/j.cmet.2016.08.001 Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 1 POMC Gene Structure and Pyrosequencing Results in an Adult Cohort (A) The POMC gene structure indicating the location of CpG islands (green), the Alu elements (black boxes) and CpGs −2 to +7, which were analyzed by pyrosequencing. (B) Bisulfite-pyrosequencing analysis of DNA methylation in peripheral blood cells from 103 normal-weight (gray) and 125 obese (red) adult individuals. The most significant differences were observed at CpG positions +3 and +5 (corresponding to nucleotide number chr2:25.384.590 and chr2:25.384.569 [UCSC human genome browser hg19]). The gray dotted line shows the variation in mean methylation values from the intron (CpG positions −2 and −1) to the exon at CpG positions +4 to +7. Error bars represent SD. (C and D) Linear regression based on the mean value of CpG positions −2 to +5 and BMI in the German adult cohort (n = 228) (C) and MSH laser-microdissected neurons from postmortem human brain samples (n = 41) (D). Cell Metabolism 2016 24, 502-509DOI: (10.1016/j.cmet.2016.08.001) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 2 Cross-Tissue Correlations in Intra-individual POMC Methylation and Stability over Time (A) POMC DNA methylation at CpG positions −2 to +5 correlates significantly between laser-microdissected MSH neurons and laser-microdissected blood cells (n = 14). (B) Intra-individual correlation between the POMC DNA methylation (%) at CpG positions −2 to +5 of postmortem human brain and kidney tissues. (C) Longitudinal stability of DNA methylation in the same individuals measured perinatally and in adolescence (n = 52). Cell Metabolism 2016 24, 502-509DOI: (10.1016/j.cmet.2016.08.001) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 3 POMC Pyrosequencing Analysis in Family Trios with One Obese Child Peripheral blood DNA samples from 47 trio families, each with one obese child, were analyzed by pyrosequencing. (A) Correlation between paternal and offspring DNA methylation at CpG positions −2 to +5. (B) Correlation between maternal and offspring DNA methylation CpG positions −2 to +5. Cell Metabolism 2016 24, 502-509DOI: (10.1016/j.cmet.2016.08.001) Copyright © 2016 Elsevier Inc. Terms and Conditions

Cell Metabolism 2016 24, 502-509DOI: (10.1016/j.cmet.2016.08.001) Copyright © 2016 Elsevier Inc. Terms and Conditions