DNMT3A mutations in AML. (A) Three conserved domains in DNMT3A are shown: the PWWP domain, which targets the enzyme to nucleic acids; the cysteine-rich PHD zinc-finger domain, which interacts with unmodified histone H3; and the highly conserved catalytic domain in the C-terminal region. The mutations in AML previously reported by us are marked in black, and the newly detected mutations in the present study are in red. The most common missense mutations are predicted to affect amino acid R882. A total of 37 AML patients had the R882H mutation, 24 with R882C, 1 with R882S, and 1 with R882P in our 1178 samples. (B) Correlation analysis of gene expression and DNA methylation. The CpG content of the promoter sequences of the genes is color coded in a separate column (left lane), including low CpG content (LCP), intermediate CpG content (ICP), and high CpG content (HCP). Hypomethylation or hypermethylation in the middle lane indicates the CpG methylated level of genes in DNMT3A-mutated samples compared with samples without DNMT3A mutations. Cluster of differently expressed genes are shown on the right. Raw microarray data of gene expression and DNA methylation were published previously.⁴⁰  (C) Quantitative RT-PCR analysis of genes associated with hematopoiesis and epigenetics regulation that were up- or down-regulated and accompanied by DNA methylation changes in microarray analysis in patients with DNMT3A mutations (DNMT3A), MLL abnormalities (MLL), or without these 2 types of aberrations (WT). (D) Quantitative RT-PCR analysis of genes in distinct HOX families in patients with DNMT3A mutations (DNMT3A), MLL abnormalities (MLL), or without these 2 types of aberrations (WT).