Fig. 4

The effects of menin and DOT1L combinatorial targeting on OC cells proliferation and transcriptome. (A) Scatter plot showing the correlation of MEN1 and DOT1L mRNA expression in OC tumors. (B) D-R Lowe graph showing the effects of combinatorial treatment with increasing MI-136 and EPZ5676 concentrations on PEO1 (left) and PEO4 (right) cells proliferation after twelve days of treatment. (C) Venn diagrams showing the number of down- (left) and up- (right) regulated transcripts in PEO1 and PEO4 cells, cultivated in the presence of 0.8 µM MI-136 and 3.2 µM EPZ5676 or vehicle (DMSO) for 9 days (|FC| ≥ Q1, padj ≤ 0.05). RNA-seq was performed in biological triplicates. (D) Heatmap and hierarchical clustering with dissimilarity measured using Manhattan distance showing common concordantly deregulated transcripts in PEO1 and PEO4 cells upon menin, DOT1L or simultaneous menin and DOT1L pharmacological inhibition with 0.8 µM MI-136, 3.2 µM EPZ5675 or 0.8 µM MI-136 and 3.2 µM EPZ5675 (|FC| ≥ Q1, padj ≤ 0.05). (E) Graphic representation of statistically significant pathways, identified by IPA analysis, in PEO1 (blue) and PEO4 (red) cells after treatment with the combination of 3.2 µM EPZ5676 and 0.8 µM MI-136 for nine days. (E) Graphic representation of determined by IPA statistically significant pathways, concordantly deregulated upon simultaneous menin and DOT1L pharmacological inhibition in both PEO1 (blue) and PEO4 (red) cells. The dashed orange line marks the B-H p-value threshold (0.05). (F) GSEA showing MYC and DNA repair GO terms highlighted by genes, deregulated upon simultaneous menin and DOT1L pharmacological inhibition in PEO1 and PEO4 cells. Negative and positive NES indicate that down- and up-regulated genes are over-represented