specific blockade of IKK activity induces the nuclear translocation of FOXO3a in leukemic cells. (A) After purification, AML blast cells were cultured during 4 hours in cytokine- and serum-free medium. During the last hour of starvation, cells were treated or not by either the anti-Nemo peptide or the peptide control. MV4-11/FOXO3a-GFP cells were cultured in 10% FCS MEM and treated or not with either the anti-Nemo peptide or the peptide control during 1 hour. Protein extracts from 10⁶ cells were analyzed by Western blot. Phospho-IKK S^176/180 was quantified and normalized to IKK signal intensity in 5 different AML samples. Results are expressed as a ratio to the control incubation without either the anti-Nemo peptide or the peptide control. The statistical significance was calculated versus the control condition (except where indicated by brackets) by a Student t test. Vertical bars indicate SDs. (B) FOXO3a localization in both AML cells from 5 different AML samples and MV4-11/FOXO3a-GFP cells treated or not with either the anti-Nemo peptide or the peptide control was investigated as described in Figure 1C. (C) For the quantification of the percentage of cells with nuclear FOXO3a, 100 cells were counted. The statistical significance was calculated versus the control condition (except where indicated by brackets) by a Student t test. Vertical bars indicate SDs. (D) Primary AML blast cells from patient 48 treated or not with the anti-Nemo peptide or the control peptide were subjected to cellular fractionation as described in Figure 1E.