Human MDS/AML cells (SKM-1 cells) were purchased from BeNa Culture Collection (Beijing, China) and cultured in Roswell Park Memorial Institute-1640 medium containing 10% fetal bovine serum in 95% humidified air with 5% CO2 at 37°C.
The small interfering RNA (siRNA)-NC-1, si-EZH1, si-NC-2, si-EZH2, si-NC-T2, and si-EHMT2 were designed and synthesized by GenePharma (Shanghai, China), and the sequence is shown inSupplementary Table 1 . Overexpression vectors of EZH2 (pcDNA3.1-EZH2) and EHMT2 (pcDNA3.1-EHMT2) and empty vectors were constructed by Zoman Biotechnology Co., Ltd. (Beijing, China). Then, the constructed vectors and siRNAs were transfected into cells using Lipofectamine 2000 (Invitrogen Inc., Carlsbad, CA, United States).
SKM-1 cells were treated with EZH2 inhibitor EPZ-6438 (5 μM, Yeasen Biotech Co., Ltd., Shanghai, China) and EHMT2 inhibitor BIX-01294 (2.5 μM, Sigma-Aldrich, Merck KGaA, Darmstadt, Germany). Briefly, cells in logarithmic growth phase were seeded into 96-well plates (1 × 105 cells/mL) supplemented with culture medium, and treated with 5 μM EPZ-6438 for 4 days (Knutson et al., 2014 (link)) and 2.5 μM BIX-01294 for 3 days, respectively (Huang et al., 2017 (link)). The treated cells were allocated as EPZ group and BIX group, respectively.
The small interfering RNA (siRNA)-NC-1, si-EZH1, si-NC-2, si-EZH2, si-NC-T2, and si-EHMT2 were designed and synthesized by GenePharma (Shanghai, China), and the sequence is shown in
SKM-1 cells were treated with EZH2 inhibitor EPZ-6438 (5 μM, Yeasen Biotech Co., Ltd., Shanghai, China) and EHMT2 inhibitor BIX-01294 (2.5 μM, Sigma-Aldrich, Merck KGaA, Darmstadt, Germany). Briefly, cells in logarithmic growth phase were seeded into 96-well plates (1 × 105 cells/mL) supplemented with culture medium, and treated with 5 μM EPZ-6438 for 4 days (Knutson et al., 2014 (link)) and 2.5 μM BIX-01294 for 3 days, respectively (Huang et al., 2017 (link)). The treated cells were allocated as EPZ group and BIX group, respectively.