The PI3K/Akt signaling pathway does not control the FOXO3a localization in AML 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 with or without IC87114. MV4-11/FOXO3a-GFP cells were cultured in 10% FCS MEM and treated with IC87114 during 1 hour. Protein extracts from 10⁶ cells were analyzed by Western blot. Phospho-Akt S⁴⁷³, FOXO3a T³², and FOXO3a S²⁵³ were quantified and normalized to Actin signal intensity in different AML samples. Results are expressed as a ratio to the control incubation without IC87114. The statistical significance was calculated by Student t test, and vertical bars indicate SDs. (B) FOXO3a localization in both AML cells from 7 different AML samples and MV4-11/FOXO3a-GFP cells treated or not with IC87114 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 by Student t test and vertical bars indicate SDs. (D) Primary AML blast cells of patient 14 treated or not with IC87114 were subjected to cellular fractionation as described in Figure 1E.