Figure 1.

MC3343 efficiently reduces DNMT protein level and enzymatic activity in EWS cells but DNA methylation status is left unaltered. (A) Western blot analysis of DNMT1 and DNMT3a, in untreated (CTRL), MC3343-treated (3–10 μM), and 5azadC-treated (3–10 μM) TC-71, A-673, LAP-35 and PDX-EW#4-C cells. Equal protein loading was monitored by anti-GAPDH blotting. One representative immunoblot of three independent experiments is shown. (B) DNMT activity measured in EWS cells after MC3343 and 5azadC treatments expressed as OD (optical density)/hours/mg (*p < 0.05; **p < 0.01, ***p < 0.001, one-way ANOVA test). The mean ± SE of three independent experiments are reported. (C) Heatmap illustrating the difference of methylation between EWS cells treated with MC3343 and 5azadC.