Effect of G9a/GLP targeting on MM cell viability, proliferation, and apoptosis. (A) Effect of UNC0638 or BIX01294 on cell viability. Cells were treated with increasing doses of UNC0638 and BIX01294. Four days later, cell viability was assessed using a CellTiter-Glo Luminescent Cell Viability Assay. Data are based on at least 3 independent experiments. (B-C) Effect of UNC0638 (UNC) or BIX01294 (BIX) on proliferation. OPM-2 and XG-20 cells were treated with the indicated concentrations of UNC0638 or BIX01294 for 48 and 72 hours. Using flow cytometry, cell cycle profiles based on DNA content were obtained after 72 hours (B), and Ki67 levels were obtained after 48 hours (C). Data are mean ± standard deviation (SD) of 3 independent experiments. (D) Effect of UNC or BIX on apoptosis. OPM-2 (left panel), XG-7 (middle panel), and XG-20 (right panel) cells were treated with the indicated doses of UNC or BIX for 48 hours. The effect on apoptosis was assessed using Annexin V-APC/7-aminoactinomycin D (7-AAD) staining, followed by flow cytometric analysis. The percentage of apoptotic cells is the sum of the percentage of Annexin V⁺ and Annexin V⁺/7-AAD⁺ cells. Data represent the mean ± SD of 3 independent experiments. (E) Effect of BIX on primary human CD138⁺ MM cells. Mononuclear cells from 12 MM patients were treated with increasing doses of BIX and cultured in the presence of IL-6 (1 ng/mL). At day 4, the percentage of viable CD138⁺ plasma cells was determined by flow cytometry. Bars represent values relative to control (cnt). *P < .05 vs control.