EZH2 inhibition induces apoptosis by upregulating the proapoptotic genes TNFSF10 and BAD. (A) SU-DHL-10 and SU-DHL-10-P cells were treated with GSK126 (10 μM) for 3 days and apoptosis was analyzed by annexin V–FITC staining. (B) TNFSF10 and BAD mRNA expression in previously published Gene Expression Omnibus (GEO) datasets including DLBCL cells treated with GSK126 (GSE: 40971). (C) TNFSF10 and BAD mRNA and protein expressions in SU-DHL-10-P and SU-DHL-10-R cells treated with GSK126 (1 μM, 48 hours). (D) TNFSF10 and BAD mRNA (bottom) or protein (top) expression in SU-DHL-10 cells expressing nonsilencing (NS) or the designated shRNAs. (E) SU-DHL-10 cells expressing nonsilencing or TNFSF10 and BAD shRNAs were analyzed using an MTT assay after treatment with the indicated concentrations of GSK126 for 72 hours. Relative proliferation under the specified conditions is shown. (F) SU-DHL-10 cells expressing the designated shRNAs were treated with vehicle or GSK126 (5 and 10 μM) for 72 hours and apoptosis was measured using an annexin V assay. (G) TNFSF10 and BAD mRNA expression in SU-DHL-10 cells expressing either PI3KCA, MEK-DD, or IGF-1R under the specified conditions. (H) FOXO3A mRNA (bottom) and protein (top) expression in SU-DHL-10 cells expressing nonsilencing or FOXO3A shRNAs. (I) TNFSF10 mRNA expression in SU-DHL-10 cells expressing nonsilencing or FOXO3A shRNAs treated with the indicated drugs for 48 hours. AUC was calculated to allow for the comparison between the 2 curves and the P values were calculated using a Student t test. Data are presented as mean ± SEM; *P < .05; **P < .01; ***P < .001.