Fig 1: Stable knockdown of RICTOR abrogated the activation of RAD001 and enhanced the inhibition of PP242 to AKT/PRAS40 signaling in vitro and in vivo. (A) ECa109 cells stably transfected with control shRNA or RICTOR shRNA were treated with RAD001 (10 μmol/L) or PP242 (2 μmol/L) for 48 h, and total proteins were extracted to analysis the expression of RICTOR, p-AKT (Ser473), AKT, p-PRAS40 (Thr246), PRAS40, p-p70S6K and p70S6K by Western blot (n = 5). (B) Total proteins in tumor tissues from nude mice were extracted and the expressions of RICTOR, p-AKT (Ser473), AKT, p-PRAS40 (Thr246), PRAS40, p-p70S6K (Thr389) and p70S6K were evaluated by Western blot (n = 5). Values represent the mean ± SD. ***P < 0.001 versus control group; ###P < 0.001 versus single-factor treatment group.