fig4

Figure 4. Cotreatment with panobinostat and CB-839 induced cytotoxicity in MM cells. (A-C) U266, RPMI 8226, and KMS-11/BTZ cells were cultured with panobinostat and/or CB-839 for 48 or 72 h. (A) Cell proliferation, (B) caspase 3/7 activity, and (C) cytotoxicity were evaluated. Statistical significance is denoted as ^****P < 0.0001 compared to the control. The experiment was repeated three times. (D) Cell cycle profiling was determined as described in the materials and method. U266 cells were treated with panobinostat (10 nM) and/or CB-839 (200 nM) for 24 h. A representative histogram for each treatment condition is illustrated. ^*P < 0.05 and ^****P < 0.0001 compared to the control. The experiment was conducted three times; (E) U266 cell lines were treated with panobinostat (10 nM) and/or CB-839 (200 nM). Total protein extracts were analyzed using immunoblotting against cleaved caspase 3, cleaved PARP, acetyl histone H4, and β-actin antibodies. MM: Multiple myeloma; RPMI: Roswell Park Memorial Institute; BTZ: bortezomib; PARP: poly (ADP-ribose) polymerase.