Fig. 5.

A germline, dominant-negative DNMT3A mutation causes focal DNA methylation alterations in human skin. (A) Whole genome bisulfite sequencing analysis for whole skin samples comparing two samples from a TBRS patient and his brother, comparing CpG methylation globally (Top) or at DMRs (Bottom). Level of significance * adjusted P < 0.05 and **P < 0.01. (B) Density plots demonstrate global CpG methylation (Top) and CpG methylation at DMRs (Bottom) in the TBRS patient and his unaffected sibling. (C) Heatmap illustrates 1,234 differentially methylated regions in the skin of the TBRS patient, compared with his sibling. (D) IGV visualization of CpG methylation at the IRX3/Irx3 and FZD1/Fzd1 loci. Each row represents an independent sample with Dnmt3a^WT vs. Dnmt3a^KO represented in blue and red (Top) and sibling vs. TBRS patient in purple and yellow (Bottom). Gray boxes highlight areas of differential methylation. (E) Gene bodies are the most frequently involved annotated region by TBRS DMRs. *There are 963 DMRs that reside in annotated gene bodies, representing 637 unique genes. (F) There are 23,983 DMRs in the Dnmt3a^KO, 16,315 of them are within 10 kb of 3,612 unique genes. Of the 1,234 TBRS DMRs, 1,086 are located within 10 kb of genes in the human genome. Of the 1,086 TBRS-DMR associated genes, 707 have a mouse homolog, with 432 genes overlapping DMR-associated genes in Dnmt3a^KO mice and the TBRS patient.