Dnmt3a loss leads to increased accessibility at enhancer chromatin marks and activation of inflammatory signaling. (A) Analysis of genome-wide differential peaks (DP) in Jak2^VF-Dnmt3a-Cas9 compared with Jak2^VF-Cas9 LSK recipient 8 weeks posttransplant. Peaks annotated according to their genomic features. Upstream or downstream ±1 kb TSS (blue), 5′ UTR (maroon), exon (orange), intron (red), 3′UTR (dark blue), ≤2 kb downstream of gene body (gray), distal intergenic (green), if not corresponding to any of previous defined feature (black). (B) DP FDR < 0.05 in Jak2^VF-Dnmt3a-Cas9 compared with Jak2^VF-Cas9 LSK recipient mice 8 weeks posttransplant within hematopoietic progenitor–specific (ST-HSCs + MPPs) enhancers, within 1 Mb of Jak2^VF-Dnmt3a-Cas9 upregulated gene. Primed (gray), active (dark gray), poised (orange) or repressed (maroon). (C) DP (FDR < 0.05), in Jak2^VF-Dnmt3a-Cas9 compared with Jak2^VF-Cas9 LSK recipient mice 8 weeks posttransplant, at promoter regions (containing H3K4Me3 marks within 1 kb up stream or downstream of TSS from nearest Dnmt3a regulated gene). Primed (gray), active (green), repressed (maroon). (D) Heat map represents DP within active enhancers. The x-axis represents the distance of the peak from the active enhancer center and each horizontal line on the y-axis represent the peaks log2 transformed fold enrichment above the background in Jak2^VF-Dnmt3a-Cas9 and Jak2^VF-Cas9 controls. (E) GSEA plot for DNMT3A-MUT pHSCs DE gene defined in human pHSCs comparing LSK from Jak2^VF-Dnmt3a-Cas9 and Jak2^VF-Cas9 recipient mice. (F) Number of DP (FDR < 0.05) within annotated enhancers (left) and annotated promoters (right) from DNMT3A-WT and DNMT3A-MUT pHSCs. Primed (gray), active (dark gray), repressed (maroon). (G) GSEA plot analyzing TNFα gene-expression signature within LSK from Jak2^VF-Dnmt3a-Cas9 and Jak2^VF-Cas9 recipient mice. NES (top) and FDR q value (bottom rows) are indicated. (H) Intracellular staining for TNFα production in LK and LSK from Jak2^VF-Dnmt3a-Cas9 and Jak2^VF-Cas9 control. TNFα staining and percentage of TNFα ⁺ (LSK or LK⁺) in Jak2^VF-Cas9 and Jak2^VF-Dnmt3a-Cas9. P values were calculated using the unpaired Student t test, *P < .05, mean ± SD. (I) GSEA plot demonstrating enrichment of TNFα gene-expression signature within MF patients with mutated DNMT3A vs nonmutated controls. (J) GSEA plot demonstrating enrichment of murine gene-expression signature derived from Jak2^VF-Dnmt3a-Cas9 LSK, within human MF with mutated DNMT3A.