Fig. 4

Mutant KMT2C results in enrichment of the MYC gene signature in prostate epithelial cells. a Schematic overview of the magnetic bead-based enrichment procedure of murine prostate epithelial cells. Sequence from left to right: 19-week-old mice were sacrificed and prostates were dissected. Tissue was enzymatically and mechanically dissociated to generate single cell suspensions. Cells were labelled with biotinylated anti-EpCAM antibody and retrieved from the bulk population using streptavidin-coated magnetic beads. The sorting efficiency was verified via flow cytometry. b Unsupervised hierarchical clustering and heatmap of significant differentially expressed genes between Pten^∆/∆ and Pten^∆/∆Kmt2c^SET∆/∆ prostate epithelial cells. 5 biological replicates were included per group. Number of genes deregulated with an adj. P value < 0.05 and log₂FC ≥ 1 / ≤ -1 are shown on the left. c HALLMARK gene sets enriched in Pten^∆/∆Kmt2c^SET∆/∆ versus Pten^∆/∆ groups at an FDR < 0.25. Dotted lines: adj. P value = -log10(0.05). d Gene expression levels of Myc based on normalized counts from RNA-Seq analysis of Pten^∆/∆ and Pten^∆/∆Kmt2c^SET∆/∆ prostate epithelial cells. Individual biological replicates are shown. Data are plotted as mean ± standard deviation, and the P value was determined by unpaired two-tailed Student’s t-test. e–h fGSEA plots of Pten^∆/∆Kmt2c^SET∆/∆ versus Pten^∆/∆ groups showing an enrichment of MYC target genes (HALLMARK_MYC_TARGETS_V1) (e), genes involved in EMT (HALLMARK_EPITHELIAL_MESENCHYMAL_TRANSITION) (f), genes upregulated in metastasis of solid tumours (RAMASWAMY_METASTASIS_UP) (g) and genes upregulated in prostate cancer lymph node metastasis versus primary prostate cancer (PCa LN Metastasis UP, see also Supplementary Materials and Methods) (h)