Enrichment of H3K79me2 on the Evi1 promoter of MLL-AF9–transformed cells. ChIP using (A) H3K79me2 and (B) H3K27me3 Abs showing enrichment of the H3K79me2 mark on the Evi1 promoter of Evi1^pos vs Evi1^neg clones. Conversely, the H3K27me3 mark is enriched on the Evi1 promoter of Evi1^neg clones compared with Evi1^posMLL-AF9 clones. ChIP using H3K4me3, H3K79me2, and H3K27me3 Abs performed on 3 EVI1^pos and 3 EVI1^neg MLL-AF9 rearranged leukemias showed higher H3K79me2 marks present on the EVI1 promoter of EVI1^pos MLL-AF9–rearranged human AMLs compared with EVI1neg cases. Q-PCRs were performed to monitor relative enrichment of each histone mark on the promoters of either (C) EVI1, (D) HoxA9 promoter, or (E) β-actin. Experiments show average relative enrichment with SD of either (A-B) 3 biologic replicates or (C-E) 2 independent ChIP pulldowns with triplicate Q-PCR measurements performed on the same cross-linked patient material.