Pharmacological inhibition of EZH2 prevents embryonal RMS cell proliferation. (a) RD cells cultured in proliferating growth medium (GM, i.e. supplemented with 10% of fetal calf serum) were treated daily with either the S-adenosyl-L-homocysteine hydrolase inhibitor 3-deazaneplanocin A (DZNep) (left panels) or the EZH2 catalytic inhibitor MC1945 (right panels) at the reported concentrations or with vehicle (i.e., water for DZNep or DMSO for MC1945) and harvested and counted at the indicated time points. *P < 0.05 (Student’s t-test); Bars, SD. Three independent experiments in duplicate. (b) Western blot showing EZH2 along with histone H3 trimethylation on Lys27 (H3K27me3), and on Lys9 (H3K9me3) levels in RD cells treated for 72 h with 5 μM DZNep (left panel) and 5 μM MC1945 (right panel) or with vehicle (i.e., water or DMSO). Total H3 and - tubulin amounts were shown as the loading controls. Representative of three independent experiments.