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  Indian J Med Microbiol
 

Figure 5: SMAD7 overexpression suppresses TGF-β1-induced SMAD2 and SMAD3 phosphorylation and nuclear translocation in fibroblasts derived from human Peyronie's disease plaque. Fibroblasts were transfected with an empty PEI25k/pCMV5 vector or a PEI25k/pCMV5-Smad7 polyplex (pSmad7) for 48 h and were then treated with TGF-β1 (10 ng ml−1) for 1 h. (a) A representative Western blot for P-Smad2, P-Smad3, and total SMAD2/3. Whole-cell extracts were fractionated in a sodium dodecylsulfate-polyacrylamide gel. (b) Data are presented as the ratio of phosphorylated protein to total protein. The relative ratio measured in the no treatment group was arbitrary presented as 1. Each bar depicts the mean values (± s.e.) from four experiments per group. *P < 0.05 by ANOVA. (c) Representative fluorescent immunocytochemistry of primary human fibroblasts with antibody against total SMAD2/3. Nuclei were labeled with the DNA dye DAPI. Schale bar = 25 ìm. (d) Nuclear fluorescence intensity was quantified for all cells. Each bar depicts the mean values (± s.e.) from four experiments per group. ***P < 0.001 by Kruskal-Wallis tests. PEI: poly (ethyleneimine); TGF-β1: transforming growth factor-β1; SMAD7: decapentaplegic homolog 7; P-Smad2: phospho-Smad2; DAPI: 4,6-diamidino-2-phenylindole; s.e.: standard error.

Figure 5: SMAD7 overexpression suppresses TGF-β1-induced SMAD2 and SMAD3 phosphorylation and nuclear translocation in fibroblasts derived from human Peyronie's disease plaque. Fibroblasts were transfected with an empty PEI25k/pCMV5 vector or a PEI25k/pCMV5-Smad7 polyplex (pSmad7) for 48 h and were then treated with TGF-β1 (10 ng ml−1) for 1 h. (a) A representative Western blot for P-Smad2, P-Smad3, and total SMAD2/3. Whole-cell extracts were fractionated in a sodium dodecylsulfate-polyacrylamide gel. (b) Data are presented as the ratio of phosphorylated protein to total protein. The relative ratio measured in the no treatment group was arbitrary presented as 1. Each bar depicts the mean values (± s.e.) from four experiments per group. *<i>P</i> < 0.05 by ANOVA. (c) Representative fluorescent immunocytochemistry of primary human fibroblasts with antibody against total SMAD2/3. Nuclei were labeled with the DNA dye DAPI. Schale bar = 25 ìm. (d) Nuclear fluorescence intensity was quantified for all cells. Each bar depicts the mean values (± s.e.) from four experiments per group. ***<i>P</i> < 0.001 by Kruskal-Wallis tests. PEI: poly (ethyleneimine); TGF-β1: transforming growth factor-β1; SMAD7: decapentaplegic homolog 7; P-Smad2: phospho-Smad2; DAPI: 4,6-diamidino-2-phenylindole; s.e.: standard error.