Figure 6.

AR is required for androgen-induced AR/Src/PI3-K/Fak complex in MDA-MB231 cells. MDA-MB231 cells were transfected with AR siRNA (a–c), using Lipofectamine^TM 2000. After transfection, the cells were made quiescent for 24 h and then left unchallenged or challenged for 5 min with 10 nM R1881, in the absence or presence of the indicated compounds. Bicalutamide (Bic; at 1μM) and the S1 peptide (at 10 nM) were added 30 min before the hormone stimulation. Cell lysates were prepared. In (a), lysates were immune-precipitated using the anti Src antibody (Anti Src Ab) or control IgG (control Ab). Proteins in immune-complexes were analyzed by Western blot, using the antibodies against the indicated proteins. In (b,c), lysates proteins were analyzed by Western blot, using the antibodies against the indicated proteins. P-Pax stands for Tyr 118-P-Paxillin, P-akt for P-Ser 473 Akt and P-Fak stands for P-Tyr397 Fak. Filters were stripped and re-probed using anti tubulin antibody, as loading control.