Fig. 5

TSP-2-induced migration was mediated through the MAPK-dependent pathway in human PCa cells. a Starved DU145 cells were incubated with TSP-2 for 10, 15, 30, 60, and 120 min, and the phosphorylation of p38, ERK, and JNK were examined by immunoblotting using anti-phospho p38, ERK, and JNK antibodies, and followed by re-probing against to total p38, ERK, JNK, and β-actin to show equal loading amounts. The DU145 cells were pretreated with the p38 inhibitor (SB203580), ERK inhibitor (U0126), and JNK inhibitor (SP600125) for 1 h (b, d, and e), transfected with p38, ERK, and JNK siRNA (c, f, and g), and stimulated with TSP-2 for 24 h. The MMP-2 mRNA expression was measured by real-time PCR (b and c), and the migratory (d and f) and invasive (e and g) abilities were assessed by the transwell analysis. Cells without treatments were used as a control (set to 100), and data were shown as multiples of that. Results are shown as the mean ± SEM (n ≥ 3, *p < 0.05 when compared to untreated control; ^# p < 0.05 when compared to the TSP-2-treated group). UT untreated control