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Figure 8. Cytochalasin D diminishes transforming growth (TGF)- –induced C-terminal Smad2 phosphorylation. Serum-starved cells were pretreated for 1 h with Cyto D before 30-min stimulation with 1 ng/ml TGF-1. Cells lysates (25 µg) were electrophoresed under reducing conditions before transfer to Immobilon-P membrane and Western analysis for C-terminal phospho-Smad2 using 1 µ/ml anti-phospho Smad2. Blots were then stripped and reprobed with 0.2 µg/ml anti-Smad 2 antibody (N19) for protein expression. Cyto D dose-dependently reduced TGF-1–simulated phosphorylation of Smad2. A representative blot is shown at the top, and the results of densitometric analysis are shown at the bottom. Values in the graph represent mean intensity of pSmad2 signal normalized to cells treated with TGF-1 along, ± SEM from five separate experiments. Degree of inhibition compared to control:¶ P < 0.020; ¶¶ P < 0.002 (Fisher PLSD).
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