Effects of refp17 and lymphoma-associated vp17 stimulation on Akt and PTEN activity in Raji cells. Cells were treated for 5 min with 0.05, 0.1, and 0.5 μg/mL of refp17 (A) and lymphoma-associated p17 variants NHL-a101 (B), NHL-a102 (C), NHL-a103 (D), NHL-a104 (E), and NHL-a105 (F). Untreated cells were used as control (lane 1). Western blot analysis of Raji lysates shows that refp17 and NHL-a103 inhibit the activation of Akt and maintain PTEN in an active state (A), as shown by the respective phosphorylation state, verified by densitometric analysis and plotting of the pAkt/Akt and pPTEN/GAPDH. On the contrary, NHL-a101, NHL-a102, NHL-a104, and NHLa105 induce the activation of Akt and maintain PTEN in an inactive state, as shown by the increased phosphorylation, verified by densitometric analysis and plotting of the pAkt/Akt and pPTEN/GAPDH. In the Upper panels, blots from one representative experiment of three with similar results are shown. In the Lower panels, values reported for phosphorylation of Akt and PTEN are the mean ± SD of three independent experiments. Statistical analysis was performed by one-way ANOVA, and the Bonferroni’s posttest was used to compare data. *P < 0.05, **P < 0.01, ***P < 0.001.