Supplementary Figure 3: Effect of inhibitor treatment on mESC properties.
From: Accurate H3K27 methylation can be established de novo by SUZ12-directed PRC2
a. Schematic representation E14 and 7d cells, which have been treated with 10 µM Ezh2 inhibitor (EPZ6438) for 7 days. b. Outline of embroid body (EB) differentiation assay used to assess differentiation capacity. c. Quantification of beating clusters following EB differentiation. E14 or 7d cells were each differentiated in both absence or presence of inhibitor. Fractions in graph show number of beating clusters over total clusters observed. d. Proliferation curve for E14 and 7d cells. 7d cells were grown with continuous presence of inhibitor. At each time point, two individual wells are counted for each cell type. This experiment was repeated as a biological replicate 2 (not shown). e. Proliferation rate (doubling time) calculated from proliferation curve in panel d and from replicate experiment 2. Error bars represent 95% confidence interval based on linear regression of proliferation curves (6 duplicate data points per curve). f. Expression level of pluripotency genes. Circles show mean value of two technical replicates for each of three biological replicates. Wide black line marks represent the mean of the biological replicates, and error bars represent the s.e.m. g. Expression level of Hoxa10, which is upregulated in Ezh1/Ezh2 dKO vs WT mESCs, in E14 and 7d cells.
