![Pharmacological inhibition of NAMPT leads to increased cytotoxicity in NAMPT-deficient cells. (A) Spearmon correlation coefficient between FK-866 AUC (GDSC2) and NAMPT mRNA expression (CCLE) in 43 cell lines of hematopoietic origin. (B) Representative western blot (left) and normalized protein densitometry analysis (right) showing NAMPT1 (guide NAMPT [gNAMPT]) clone 1 [mean, 0.45; standard deviation (SD), 0.072] and gNAMPT clone 2 [mean, 0.2507; SD, 0.11]) and β-ACTIN protein levels in control (HPRT) and NAMPT-deficient clones (n = 2 clones per genotype; n = 3 replicates per clone) of the OCI-AML3 cell line, which is triploid for NAMPT. (C) Representative dose-response curves of OCI-AML3 control (HPRT) and NAMPT-deficient clonal cell lines treated with the KPT-9274 for 24 hours. (D) Average AUC and IC50 values of OCI-AML3 control (HPRT) and NAMPT-deficient clonal cell lines treated with the KPT-9274 for 24 hours (n = 7). (E) Annexin V/7-ADD flow cytometric analysis of cell death of OCI-AML3 control (HPRT) and NAMPT-deficient clonal cell lines treated with 25-nM KPT-9274 for 48 hours (n = 3). (F) Annexin V/7-AAD flow cytometric analysis of cell death of OCI-AML3 control (HPRT) and NAMPT-deficient cells treated with 25-nM KPT-9274 for 48 hours in the presence or absence of Z-VAD-FMK (50 μM), Fer-1 (1 μM), or Nec-1 (10 μM). Data were normalized to 0-nM (dimethyl sulfoxide [DMSO]) condition of each genotype. One-way ANOVA with Dunnett or Sidak correction for multiple comparison tests was used for statistical analysis among 3 independent groups. AUC, area under the curve; Fer-1, ferrostatin-1; TPM, transcripts per million.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodneoplasia/2/3/10.1016_j.bneo.2025.100119/1/bneo_neo-2024-000529-gr3.jpeg?Expires=1757529162&Signature=nO2em7su7Ix~WoUHSVsmkKiJvMrj~RRa4ylCKFTzBe4qVjUqt4faDFQM83txUpO-eYcn1mdBArMaPLEi8GziCNv5wzHLyC~gzy65vhKCIrCpdaYNj9pQhm4brNzwwrpX04B89nMjbtUbSgkVU4T~k7mMIZljXXupBeL3n5kuCubZyTx2paxasflgAYC9eHE9fdS5tMgxwNh~dHJOvxNUHxqICxKk-AYervIrnNuXhGAUBxakHpV7wQ7xcSXVPzL1955ssH~2~l32ovx0uNRtmDn8r6seq5fiPFHY3LZvS-BawEQun58SaIMLcZvyJ8QZi0r9qkO2eKm6A3zdcZbn5A__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Pharmacological inhibition of NAMPT leads to increased cytotoxicity in NAMPT-deficient cells. (A) Spearmon correlation coefficient between FK-866 AUC (GDSC2) and NAMPT mRNA expression (CCLE) in 43 cell lines of hematopoietic origin. (B) Representative western blot (left) and normalized protein densitometry analysis (right) showing NAMPT1 (guide NAMPT [gNAMPT]) clone 1 [mean, 0.45; standard deviation (SD), 0.072] and gNAMPT clone 2 [mean, 0.2507; SD, 0.11]) and β-ACTIN protein levels in control (HPRT) and NAMPT-deficient clones (n = 2 clones per genotype; n = 3 replicates per clone) of the OCI-AML3 cell line, which is triploid for NAMPT. (C) Representative dose-response curves of OCI-AML3 control (HPRT) and NAMPT-deficient clonal cell lines treated with the KPT-9274 for 24 hours. (D) Average AUC and IC50 values of OCI-AML3 control (HPRT) and NAMPT-deficient clonal cell lines treated with the KPT-9274 for 24 hours (n = 7). (E) Annexin V/7-ADD flow cytometric analysis of cell death of OCI-AML3 control (HPRT) and NAMPT-deficient clonal cell lines treated with 25-nM KPT-9274 for 48 hours (n = 3). (F) Annexin V/7-AAD flow cytometric analysis of cell death of OCI-AML3 control (HPRT) and NAMPT-deficient cells treated with 25-nM KPT-9274 for 48 hours in the presence or absence of Z-VAD-FMK (50 μM), Fer-1 (1 μM), or Nec-1 (10 μM). Data were normalized to 0-nM (dimethyl sulfoxide [DMSO]) condition of each genotype. One-way ANOVA with Dunnett or Sidak correction for multiple comparison tests was used for statistical analysis among 3 independent groups. AUC, area under the curve; Fer-1, ferrostatin-1; TPM, transcripts per million.
