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. 2012 Jul 22;33(11):2054–2064. doi: 10.1093/carcin/bgs252

Fig. 2.

Fig. 2.

Activation of Nodal and TGF-β signaling in prostate cell lines. (A) Western blot analyses of phosphorylated Smad2 and Smad3 and β-actin in PZ-HPV7, DU145 and PC3 cells at different time periods after treatment with Nodal (200ng/ml) or TGF-β1 (5ng/ml). Western blots using anti-β-actin antibody were used as internal controls. Quantitative analysis of p-Smad2 and p-Smad3 in PZ-HPV7, DU145 and PC3 cells treated with Nodal and TGF-β were relative to that of the untreated control (designated as one) after normalization to the signal obtained with Smad2/3. Each bar represents mean ± SEM (n = 3). *Significantly different (P < 0.05) compared with untreated controls. (B) Western blot analyses of phosphorylated Smad2 and Smad3, total Smad2/3 and β-actin in PC3 cells after pretreatment with Smad3 inhibitor (SIS3, 1 µM). (C) Pretreatment with SIS3 (1 µM) for 30min completely blocked the migration of PC3 cells induced by TGF-β (5ng/ml) but not in Nodal (500ng/ml) and EGF (3ng/ml) treated cells. The data were presented as mean ± SEM (n = 3). *Significantly different (P < 0.05) compared with untreated controls.