Cytotoxic effects of KPT-9274 are independent of TP53 mutational status in cells with NAMPT deficiency. (A) Venn diagram showing TP53 mutational or deletion status in cases of −7/del(7q) NAMPT haploinsufficient patients from the TCGA data set. (B) Western blot showing NAMPT, TP53, β-ACTIN, and lamin protein levels in control (HPRT), TP53-null, NAMPT-deficient, TP53-null, and TP53-null and NAMPT-deficient clonal cell lines of OCI-AML3. (C) Representative dose-response curves (left) and AUC and IC₅₀ graphs (right) of control, TP53-null, NAMPT-deficient, and TP53-null NAMPT-deficient clonal OCI-AML3 cell lines treated with the KPT-9274 for 24 hours (n = 4). (D) Annexin V/7-ADD flow cytometric analysis of cell death control, TP53-null, NAMPT-deficient, and TP53-null NAMPT-deficient clonal OCI-AML3 cell lines treated with 25 nM KPT-9274 for 48 hours (n = 3). (E) Western blot showing cleaved caspase 3 and β-ACTIN levels in control (HPRT), TP53-null, NAMPT-deficient, and TP53-null NAMPT-deficient OCI-AML3 whole-cell extracts. Cells were treated with 25-nM KPT-9274 for 24 and 48 hours. A control (HPRT) cell line treated with 1-μM staurosporine for 6 hours was used as a positive control for apoptosis. One-way ANOVA with Dunnett correction for multiple comparison tests were performed for statistical analysis among 4 independent groups.