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Fig. 7. The effect of p38 inhibitors MAPK and PI3K on Smad7-dependent activation of the actin filament system. (A) PC-3U/pMEP4-S7 cells were starved in 1% FBS for 12 hours and pretreated for 1 hour with 10 µM SB203580, before the addition of 1 µM of CdCl₂ for 12 hours or 24 hours, or 1 µM CdCl₂ for 12 hours together with 30 minutes, 12 hours or 24 hours of TGF-ß. Flag-tagged Smad7 was visualized by a Flag-specific antibody followed by an FITC-labeled anti-mouse antibody. Filamentous actin was visualized by TRITC-labeled phalloidin. Arrowheads indicate cells with stress fibers (24 hours, upper panel) and cells lacking stress fibers (24 hours, lower panel). Bars, 20 µm. (B) PC-3U/pMEP4-S7 cells were transiently transfected with dominant negative mutant p38 MAPK and starved in 1% FBS for 12 hours. The cells were treated with 1 µM CdCl₂ for 12 hours. The expression of dominant negative p38 MAPK was visualized with a mouse monoclonal anti-p38 MAPK antibody followed by a TRITC-labeled anti-mouse antibody. Smad7 was detected by a goat anti-Smad7 antibody followed by an Alexa Fluor 488-labeled anti-goat antibody. Arrowheads denote transfected cells. Bar, 20 µm.

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