Activation of PI3K and Akt pathway and its role in FMRP production and cell viability. ETO-treated HeLa cells were analyzed using Western blot to investigate the activation of PI3K (A) and Akt (B). Western blot against total protein was used as a loading control. (C) An inhibitor of Akt phosphorylation, LY294002 (10 μM, LY) was pretreated and the level of FMRP was determined by Western blot. (D) After LY treatment, cell viability was measured by MTT assay, inhibition of Akt phosphorylation further decreased cell viability. (E) The level of BcL-xL after LY treatment. Increased BcL-xL expression induced by etoposide treatment was prevented by LY294002 treatment. (F) Another inhibitor of Akt, inhibitor VIII (0, 5, 10 μM, VIII) pretreatment also decreased activity and expression of Akt and Bcl-xL, respectively, in a concentration dependent manner. At the same time, cellular viability was also reduced by VIII treatment. The bar graphs represent the quantification of band intensity. Data represent mean ± S.E.M. * significantly different as compared with control and # significantly different as compared with ETO alone treated sample (p < 0.01, n = 4).