Presence of Gfi136N induces epigenetic changes at several loci. Result of the ChIP-Seq experiments indicating Gfi1 occupancy and histone H3 lysine 4 9H3K4) at selected gene loci. For the detection of H3K4 dimethylation, ChIP was performed with sorted Lin⁻, c-kit⁺, Sca1⁻ cells from the indicated mouse strains using a α-H3K4 dimethyl antibody. For the detection of Gfi1, ChIP was performed with an α-Gfi1 antibody using MLL-ENL-transduced AML cells. After immunoprecipitation, DNA-protein complexes were subjected to high throughput sequencing to determine the genome-wide distribution of H3K4 dimethyl as well as of Gfi1 binding. In each figure, the top half represents the enrichment for Gfi1 binding and the lower half the distribution of H3K4dimethylation in cells from either Gfi1^36N/36N or Gfi1^36S/36S mice. The results of the H3K4dimethyl ChIP-Seq were normalized (see “ChIP seq”), and the curves of obtained with cells from the 2 different mouse strains were superimposed. The curves representing Gfi1^36N/336N cells appear in gray, the curves of Gfi1^36S/36S in black. The following gene loci have been analyzed: Hoxa9 (A), Id2 (B), Meis1 (C), IL-6Ra (D), PU.1 (E), and Pbx1 (F). (G) Summary of changes in H3K4 dimethylation and expression of the indicated Gfi1 target and control genes. The fold change in H3K4dimethylation between cells from Gfi1^36N/36N and Gfi1^36S/36S mice was calculated by dividing the areas under the respective curves. The area under the curves was determined using the TL-100 Version 2008 software from Non-Linear Dynamics. Fold change expression of the indicated genes between cells from Gfi1^36N/36N and Gfi1^36S/36S mice was calculated by dividing the absolute expression levels obtained from the DNA microarray expression analysis (Figure 2D).