Fig. 7

BCAT1-induced aggressive behaviors of CRPC cells are dependent on the activation of ERK5 signalling. A Intracellular glutamate levels were determined in BCAT1-overexpressing PC3 and DU145 cells. B The protein expression levels of ERK5, p-ERK5, ERK1/2, p-ERK1/2, p38, p-p38, JNK, and p-JNK in BCAT1-overexpressing PC3 and DU145 cells were determined by western blotting. The expression intensity was normalized to that of β-tubulin (right). C PC3 and DU145 cells were treated with 10 μM glutamate for 24 h, after which the protein expression levels were analysed by western blotting. The expression intensity was normalized to that of β-tubulin (right panel). D BCAT1-overexpressing cells were treated with JNJ16259685 (80 μM) for 24 h, after which the protein expression levels were analysed by western blotting. The expression intensity was normalized to that of β-tubulin (right panel). E–G BCAT1-overexpressing cells were treated with JNJ16259685 (80 μM) for 24 h, after which proliferation (E), colony formation (F), migration, and invasion (G) were measured. The right show the quantitative results. H Representative immunoblots of HuR, BCAT1, ERK5, and p-ERK5 in HuR KO1 PC3 and HuR KO4 DU145 cells with or without BCAT1 overexpression and incubation with JNJ16259685 (80 μM) for 24 h (left). The right show the expression intensity normalized to that of β-tubulin. p values are shown for each comparison (^**p < 0.01, ^#p < 0.05, ^##p < 0.01, ^&p < 0.05, ^&&p < 0.01)