Figure 2.

Effects of DZNep on PRC2 proteins and histone methylations. (A) MCF-7 and HCT116 cells were treated with 5 μM DZNep for 48 and 72 h and cells were harvested for Western blot analysis using antibodies detecting the indicated proteins. (B) MCF-7 cells were treated with DZNep for 48 h and the levels of indicated histone methylations were examined by Western blot analysis. (C) MCF-7 cells were treated with DZNep for the indicated times and cells were harvested for Western blot analysis of the indicated proteins. (D) MCF-7 and HCT116 cells were treated as in A and total RNA was isolated for RT–PCR analysis of EZH2, EED, and SUZ12 mRNA levels. (E) MCF-7 cells were treated with DZNep for 18 h, followed by the addition of proteosome inhibitors MG132 (5 μM), LLNL (50 μM), or MG115 (20 μM) for 8 h. Cells were harvested for Western blot analysis of EZH2 and SUZ12. (F) MCF-7 cells were treated with siRNA targeting EZH2, EED, or SUZ12 individually. After 72 h, cells were harvested and the levels of PRC2 proteins and H3-K27me3, H3-K9me3, and total H3 were examined by Western blot analysis as indicated. (G) MCF-7 and MCF-10A cells were treated as in F and cell death was measured by PI staining and FACS analysis. Data represent ±SD from three independent experiments.