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. 2021 Jan 29;22(3):1341. doi: 10.3390/ijms22031341

Figure 5.

Figure 5

Cotreatment with CFZ and A452 leads to synergistic apoptosis induction in both BTZ-sensitive and BTZ-resistant U266 MM cells. (A) BTZ-sensitive U266 and BTZ-resistant U266 (U266/VelR-1 and U266/VelR-2) MM cells were treated with 0.1% DMSO (control), A452 (1 μM), and carfilzomib (CFZ) (100 nM) or in combination with these compounds as indicated for 24 h. Apoptotic markers were identified by immunoblotting using whole-cell lysates. Glyceraldehyde 3-phosphate dehydrogenase (GADPH) and α-tubulin were used as equal loading controls. The abundance of the indicated proteins was semi-quantified relative to GAPDH or α-tub; control levels were set at 1. (B) BTZ-sensitive U266 and (C) BTZ-resistant U266/VelR-1 MM cells were treated with 0.1% DMSO (control), A452 (1 μM), and CFZ (100 nM) or in combination with these compounds as indicated for 36 h. Cell death was assessed by flow cytometry and Annexin V/PI staining. Data present as mean ± SD (n = 3). * p < 0.05, ** p < 0.01, and *** p < 0.001 vs. DMSO control; ## p < 0.01 and ### p < 0.001 vs. A452-treated group; $$ p < 0.01 vs. CFZ-treated group, two-way analysis of variance (ANOVA) test.