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Figure 1


Fig. 1. Effects of cholesterol and LDL on Smad2 phosphorylation (A-C) and nuclear translocations (D) in Mv1Lu cells and BAECs stimulated with TGF-beta1. Cells were treated with increasing concentrations of cholesterol, as indicated (A,B), 50 µg protein/ml LDL (C), 5 µg protein/ml VLDL (C) or 50 µg/ml cholesterol (D) at 37°C for 1 hour and then further incubated with 50 pM TGF-beta1 for 30 minutes. P-Smad2 and total Smad2 in the cell lysates were analyzed by immunoblotting. The relative level of P-Smad2 (P-Smad2/Smad2) was estimated. A representative of a total of three analyses is shown (top). The quantitative analysis of the immunoblots is shown below. The relative level of P-Smad2 in cells treated with TGF-beta1 only was taken as 100% of TGF-beta1-stimulated Smad2 phosphorylation. The data are mean ± s.d. *,**Significantly lower than that in cells treated with TGF-beta1 only: P<0.001 and P<0.05, respectively. (D) Smad2 nuclear translocation was analyzed by indirect immunofluorescent staining. Rhodamine fluorescence represents P-Smad2 staining (a-c) whereas the nuclei were stained by DAPI staining (d-f).





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