Figure 6.

PRMT5-mediated repression of H3K27 tri-methylation contributes to its cell-cycle effects. (A) Several leukemia cell lines and 2 AML patient samples were treated with different concentration of PRMT5i or EZH2i alone, or different combination of these two inhibitors for 6 days. Cell viability was determined by Celltiter-Glo assay. The synergy of these two inhibitors was determined by Bliss model using the Synergyfinder software. Negative synergy scores represent overall antagonizing effect of these two inhibitors; while positive scores mean synergistic effect. (B) Molm13 cells were treated with DMSO, EZH2i, PRMT5i and the combination of these two inhibitors, cells were counted at day 4 and day 6. n = 4. (C) Molm13 cells were treated with DMSO, PRMT5/EZH2 inhibitors for 4 days. Cells were fixed in ethanol and subjected to PI staining for cell-cycle analysis. Percentage of cells in S and G2/M phase was plotted here. n = 4. (D). A representative FACS plot of cell cycle is shown here. Ei: EZH2 inhibitor; P5i: PRMT5 inhibitor; Numbers on the X-axis indicate the concentration of PRMT5 inhibitor in uM; and EZH2 inhibitor was used at 1 µM in Figure 6B–D.