a, Bottom, ENL protein structure, with the region that is mutated in cancer shown in red. Above, amino-acid sequences of the T1 to T8 tumour-associated mutations and the corresponding WT region. b, c, Western blots showing the levels of ectopically expressed WT or mutant Flag–ENL proteins in HEK293 (b) and HK-2 (c) cells. Independent experiments were repeated four times with similar results. β-Tubulin is used as a loading control. d, e, Venn diagrams showing the number and overlap of genes for which expression is significantly upregulated upon expression of mutant ENL as compared with WT ENL in HEK293 (d) and HK-2 (e) cells. Genes with a fold change of 1.5 or more and a false discovery rate (FDR) of 0.01 or less are considered to be significantly upregulated. f, g, GSEA plots evaluating the changes in the indicated gene signatures upon expression of the indicated ENL mutants compared with WT in HEK293 (f) and HK-2 (g) cells. h, i, Volcano plots of RNA-sequencing data demonstrating the −log 10 P-values versus log 2 fold changes in HEK293 (h) and HK-2 (i) cells. HOXA genes are highlighted in red. P-values were determined by two-tailed exact test, adjusted by FDR. j, Western blot showing the close-to-endogenous levels of ectopically expressed WT or mutant Flag–ENL in HEK293 cells. Independent experiments were repeated three times with similar results. k, mRNA expression analysis (normalized to GAPDH) of selected ENL target genes in HEK293 cells (from panel j) expressing the indicated constructs. vec, vector control. Data represent mean ± s.e.m., n = 3 technical replicates, independent experiments were repeated three times with similar results. l, western blot showing the protein levels of ectopically expressed wildtype or indicated mutants (as illustrated in a) Flag-ENL in HEK293 cells. Experiment repeated three times independently with similar results. m, mRNA expression analysis (normalized to GAPDH) of selected ENL target genes in HEK293 cells (from panel l) expressing the indicated constructs. Vec, vector control. Data represent means from n = 2 technical replicates; results are representative of three independent experiments. For gel source data (b, c, j, l), see Supplementary Fig. 1. Source data