Clinical Validation of a Genome-Wide DNA Methylation Assay for Molecular Diagnosis of Imprinting Disorders  Erfan Aref-Eshghi, Laila C. Schenkel, Hanxin Lin, Cindy Skinner, Peter Ainsworth, Guillaume Paré, Victoria Siu, David Rodenhiser, Charles Schwartz, Bekim Sadikovic  The Journal of Molecular Diagnostics  Volume 19, Issue 6, Pages 848-856 (November 2017) DOI: 10.1016/j.jmoldx.2017.07.002 Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Methylation changes in imprinting regions of patients with Angleman syndrome (AS), Prader-Willi syndrome (PWS), and Beckwith-Wiedemann syndrome (BWS). A: Hypermethylation and hypomethylation of SNRPN 5′ untranslated region (UTR) in patients with PWS (red lines) and AS (green lines), respectively, compared with healthy controls (purple lines) (methylation levels for multiple CpG sites from individuals from the healthy population). B: Hypomethylation of KCN11QT1/KCNQ1 among patients with BWS (red lines) compared with healthy controls (purple lines). The Journal of Molecular Diagnostics 2017 19, 848-856DOI: (10.1016/j.jmoldx.2017.07.002) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 Methylation changes in three imprinted regions in chromosome 7 in a single patient with Silver-Russell syndrome (SRS). The figure shows hypermethylation in the GRB10 (A), MEST (B), and PEG10 (C) genes in one patient with SRS (teal lines) compared with healthy controls (purple lines). Gray rectangles indicate CpG islands. The Journal of Molecular Diagnostics 2017 19, 848-856DOI: (10.1016/j.jmoldx.2017.07.002) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 Hierarchical clustering of patients with imprinting disorders and controls. Rows represent CpG probes from the disease-specific imprinted regions; column, samples; dark blue, higher methylation; light blue/white, lower methylation (β value range, 0 to 1). A: Patients with Prader-Willi syndrome (blue bar) have significant hypermethylation, whereas those with Angelman syndrome (red bar) have loss of methylation in the seven probes illustrated here compared with the healthy controls (green bar). B: Patients with Beckwith-Wiedemann syndrome (red bar) have methylation loss in the 34 probes from the imprinted region compared with the controls (blue bar). C: One patient with Silver-Russell syndrome (red bar) represents a significant gain of methylation among all the probes overlapping three imprinted segments in chromosome 7 compared with controls (blue bar). The Journal of Molecular Diagnostics 2017 19, 848-856DOI: (10.1016/j.jmoldx.2017.07.002) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Supplemental Figure S1 Hypermethylation of Mcts2/HM13 in two patients with intellectual disability (red and blue lines). Gray rectangle indicates CpG island; gray lines, methylation levels for multiple CpG sites in the healthy population. The Journal of Molecular Diagnostics 2017 19, 848-856DOI: (10.1016/j.jmoldx.2017.07.002) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Supplemental Figure S2 Hypomethylation at a CpG island at the promoter of FAM50B in two patients with intellectual disability (red and blue lines). Gray rectangle indicates CpG island; gray lines, methylation levels for multiple CpG sites in the healthy population. The Journal of Molecular Diagnostics 2017 19, 848-856DOI: (10.1016/j.jmoldx.2017.07.002) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Supplemental Figure S3 Hypomethylation of ZNF597 promoter in one patient previously diagnosed with Coffin-Siris syndrome (red line). Gray rectangle indicates CpG island; gray lines, methylation levels for multiple CpG sites in the healthy population. The Journal of Molecular Diagnostics 2017 19, 848-856DOI: (10.1016/j.jmoldx.2017.07.002) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions