Figure 8.

Tropomyosin receptor kinase A (TrkA) activation by Nerve growth factor (NGF) triggers mitogen-activated protein kinases (MAPKs) and Akt activation in castration-resistant prostate cancer (CRPC) cells. Quiescent C4-2B (A, E and H), DU145 (B, F and I) and PC3 (C, G and L) cells were used. In A–C, cells were left un-stimulated or stimulated for the indicated times with 100 ng/mL NGF. In D, the indicated cycling cells were used. In E–L, cells were left un-stimulated or stimulated for the indicated times with 100 ng/mL NGF, in the absence or presence of GW441756 (GW; 1μM). In A-C, protein lysates were analyzed by western blot, using the indicated antibodies. Filters were re-probed using anti-ERK (Extracellular signal-regulated kinase) or anti-Akt or anti-tubulin (tub) antibodies, as a loading control. In (D), protein lysates were analyzed by Western blot, using the indicated antibodies. In E–F, protein lysates were analyzed by western blot, using the antibodies against the indicated proteins (p-TrkA stands for P-Tyr-490 TrkA). In H–L, cells were left untreated (untr) or treated for the indicated times with NGF (100 ng/mL), in the absence or presence of 1μM GW441756 as reported in E–G. Lysates proteins were analyzed by western blot, using the antibodies against the indicated proteins. In A, B, C, H, I, L, p-ERK stands for P-Tyr 204 ERK 1, and the corresponding phosphorylated ERK 2; p-Akt stands for P-Ser 473 Akt. The filters were re-probed using anti-ERK or anti-Akt or anti-tubulin (tub) antibodies, as loading controls.