CRM1 inhibition induces differentiation of AML cell lines. (A) CD11b measurement by FACS in AML cell lines after KPT-185 treatment at 72 hours. (B) Giemsa stain of cytospins of Kasumi-1 and MV4-11 cells treated with KPT-185 or controls. Magnification, 40×. Arrows indicate nuclear condensation. (C) CEBPA protein expression in MV4-11 cells and OCI-AML3 cells after KPT-185 treatment or controls for 24 and 48 hours. Loading control is actin. (D) GCSFR and lysozyme mRNA expression after KPT-185 treatment or controls in MV4-11 cells and OCI-AML3 cells. Results are shown as fold change after normalization with 18s and 2ΔC_t calculations. (E) CEBPA protein expression after p53 siRNA or scramble oligonucleotide transfection in OCI-AML3 cells subsequently treated with KPT-185 or controls for 24 hours. Protein expression of p21 was measured as control for successful p53 inhibition. (F) CD11b expression in OCI-AML3 cells after transfection with scramble or p53 antisense oligonucleotides and treatment with DMSO or KPT-185 at 72 hours. (G) Apoptosis as measured by annexin V/PI staining using FACS at 48 hours after scramble or p53 antisense oligonucleotides and treatment with DMSO or KPT-185.