MMSET overexpression alters normal H3K36me2 distribution.

<p>(A) Loss of MMSET expression in t(4;14)+ cells depletes H3K36 methylation and increases H3K27 methylation. Immunoblot of nuclear extracts from t(4;14)+ KMS11 cells with inactivated translocated (TKO) or wild-type (NTKO) <i>MMSET</i> allele were probed using antibodies for MMSET, H3K36me2, H3K27me3 and total H4 as control. (B) Heatmaps of H3K36me2 distribution between NTKO (left) and TKO (right) cells. On the y-axis, genes were ranked based on their expression level from high (top) to low (bottom) and on the x-axis, normalized read density from ChIP-seq experiments was plotted in gene bodies and 10 kb upstream and downstream from the transcription start (TSS) and end (TES) sites. (C) MMSET alters H3K36me2 distribution in intragenic (top) and intergenic (bottom) regions. (Top) Average read density across 15,386 genes in NTKO (left, red) and TKO (right, green) cells. (Middle) UCSC genome browser view of the H3K36me2 distribution on a representative locus (<i>BTF3</i>) in NTKO and TKO cells. (Bottom) Average normalized tag density of H3K36me2 in 6,172 intergenic regions in NTKO (left) and TKO (right) cells. (D) Representative UCSC genome browser view of the H3K36me2 distribution on chromosome 8 of NTKO (red) and TKO (green) cells. Top panel encompasses a gene-rich region of the chromosome and the bottom insert represents a 1MB region of the 8q24 gene desert. (E) ChIP-qPCR for H3K36me2 on the <i>BTF3</i> locus. Methylation enrichment was tested 5 kb upstream of the TSS (5′) and at the TSS (right). Two independent biological replicates are shown.