<p>(A) HeLa cells were transfected with siRNA directed against RICTOR and then allowed to adhere to plates coated with invasin (β1 integrin-ligand) or Pluronic (non-adhesive control) for 60 min. Cell lysates were subjected to SDS-PAGE (4–15% gradient gel) followed by western blotting using the different antibodies as indicated. A representative western blot is shown and quantifications of AKT pSer473, FOXO1 pThr24, and BAD pSer136 on Pluronic and invasin are given below (mean ± s.e.m.; <i>n</i> = 3; * represents <i>p</i><0.005 and ** represents <i>p</i><0.05). (B) MCF7 cells were transfected and treated in the same manner as in (A). (C) HeLa cells transfected with ILK-directed siRNA or non-target siRNA were allowed to adhere to plates coated with Pluronic, collagen type I or invasin. Cells were lysed and analysed as explained above. A representative western blot is shown and the graph to the right shows quantification of AKT pSer473 levels on Pluronic and invasin (mean ± s.e.m.; <i>n</i> = 3). (D) HeLa cells were transfected simultaneously with PAK1- and PAK2-directed siRNAs or non-target siRNA. Adhesion assays were performed on plates coated with Pluronic, invasin or collagen type I. Cell lysates were subjected to SDS-PAGE (10% gel) followed by western blotting using the different antibodies as indicated. A representative western blot is shown and the graph below presents quantification of AKT pSer473 levels on Pluronic and invasin (mean ± s.e.m.; <i>n</i> = 3). (E) Adhesion assay with MCF7 cells transfected with PAK1- and PAK2-directed siRNAs or non-target siRNA, performed as described in (A).</p
