Fig. 5.

EZH2 inhibition is associated with NK cell activation. a Gene set enrichment analysis shows that EZH2 inhibition alone or in combination with cisplatin is associated with increased in IFN-γ signaling. qPCR analysis showed that NK cell activity associated transcripts IFN-γ, MIP-1α, CD3D, CYTH1, CD86, ICAM1, and ICAM2 were significantly upregulated in the combination treatment as compared to cisplatin alone treatment. Error bars represent standard error of means of all the mice in each group. One-way ANOVA model with pairwise comparisons made as appropriate were performed to test differences between single and combination treatments. *p < 0.05, **p < 0.01, triangle indicates significant difference as compared to vehicle, hash indicates significant difference between the EPZ011989 and cisplatin alone treatments. b, c CD56, an NK cell-associated marker is significantly higher in combination-treated tumors compared to single agent and vehicle-treated mice. NCR1, Natural Cytotoxic triggering Receptor 1, that mediates tumor cell lysis is significantly increased in the EPZ011989 and combination-treated groups compared to vehicle and cisplatin-treated groups. Orange arrows indicate infiltrating cells and black arrows point to tumor cells. Error bars represent standard error of means of Image J quantitation in all the mice in each group. One-way ANOVA model with pairwise comparisons made as appropriate were performed to test differences between single and combination treatments. *p < 0.05, **p < 0.01, triangle indicates significant difference as compared to vehicle, hash indicates significant difference between the EPZ011989 and cisplatin alone treatments. d RNA-sequencing of a 66 tumors revealed significant alterations in gene expression of a subset of ligands and receptors that are involved in the activation of natural killer (NK) cells. e z-scores of NK related transcripts are higher in tumors with no mutations compared to tumors with KDM6A and/or SWI/SNF mutations