Effect of BIX01294 treatment on tumor progression in the murine 5TGM1 model. Effect of G9a/GLP targeting on murine MM cell viability (A) and apoptosis (B). (A) The murine MM cell lines 5T33vv, 5T33vt, and 5TGM1 were treated with increasing doses of UNC0638 (UNC) and BIX01294 (BIX). After 4 days, cell viability was assessed using a CellTiter-Glo Luminescent Cell Viability Assay. Data are based on ≥3 independent experiments. (B) Effect of BIX01294 on apoptosis. 5T33vt and 5TGM1 cells were treated with the indicated doses of BIX01294 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 are mean ± standard deviation of 3 independent experiments. (C-E) Effect of G9a/GLP targeting on tumor burden and OS in the murine 5TGM1 model. (C) Scheme depicting the experimental setup. Mice were inoculated with 5TGM1 cells and treated for 14 days with vehicle (n = 11) or with 10 mg/kg BIX01294 (n = 11) 3 times a week. Treatment started 14 days after tumor inoculation. (D) Tumor burden was monitored at day 30 by BLI measuring total photon flux. BLI images from 3 mice from each treatment group are shown. (E) Effect of BIX01294 on OS. Difference in OS was assayed with a log-rank test, and survival curves were plotted using the Kaplan-Meier method. *P < .05.