Fig. 2: DOT1L mediates cell competition and gene expression.
From: The histone-methyltransferase DOT1L cooperates with LSD1 to control cell division in blast-phase MPN
A Illustration of the experimental strategy to isolate DOT1L-ko clones. B Western blot showing global H3K79me2 abundance in histone extracts (right) or DOT1L protein levels in nuclear extracts (left) from DOT1L-ko HEL clones with partial or total loss of DOT1L activity (sgDOT1L-clone 1; sgDOT1L-clone 2). C Bar graphs showing the results of a fluorochrome-based cell competition assay. Relative abundance of empty vector control or DOT1L-sgRNA expressing HEL cells as compared to non-transduced competitor cells in a chimeric mixture are shown. D Kaplan–Meyer curves showing survival of NXG mice engrafted with 50,000 HEL cells harboring a complete DOT1L-ko (clone 2) or empty vector control. Pie charts show the fraction of sgRNA vector expressing cells in human CD45+ cells in the bone marrow of mice that developed disease, indicating counterselection of the BFP+ cells in the animals of the DOT1L-ko cohort. Statistics: log-rank test, ***p < 0.0001. E Volcano-plot showing differentially expressed genes (DEGs) from RNAseq in HEL cells after knockout of DOT1L (clone 2). F Geneset-enrichment analysis plots from RNAseq in HEL cells after knockout of DOT1L. G Tornado-plots visualizing the global occupancy of DOT1L and deposition of H3K79me2 across all transcription start sites (TSS) in ChIPseq of HEL cells. H Heatmap showing the fractions of UP- and DOWN-regulated DEGs after DOT1L-ko among those genes associated with the top 3000 DOT1L-bound TSS. Selected genes from the downregulated cluster are annotated.
