Journal: The World Journal of Men's Health

Article Title: Vactosertib, a Novel, Orally Bioavailable Activin Receptor-Like Kinase 5 Inhibitor, Promotes Regression of Fibrotic Plaques in a Rat Model of Peyronie's Disease

doi: 10.5534/wjmh.190071

Figure Lengend Snippet: Vactosertib suppresses Smad2 and Smad3 phosphorylation and nuclear translocation. Fibroblasts were pretreated for 24 hours with Vactosertib (400 ng/mL) then exposed with transforming growth factor-β (TGF-β1) (10 ng/mL) for 1 hour. (A) Representative fluorescent immunocytochemistry of primary human fibroblasts cells stained with antibodies against total Smad2/3. Nuclei was dyed using DNA dye, DAPI (4,6-diamidino-2-phenylindole). Scale bar indicates 100 µm. (B) Nuclear fluorescence signal was quantified for all cells. Each bar depicts the mean value (±standard errors) of four experiments per group. a p<0.05, compared with the control group; b p<0.05, compared with the TGF-β1 group. (C) Representative Western blot analysis for phosphorylated Smad2 (p-Smad2), phosphorylated Smad3 (p-Smad3), total Smad2/3, phospho-p38 (p-p38), and p38. (D) Data are expressed as the ratio of phosphorylated protein/total protein. The data measured relative to the control group ratio is arbitrarily presented as 1. Each bar depicts the mean value (±standard errors) of four experiments per group. a p<0.05, compared with the control group; b p<0.05, compared with the TGF-β1 group.

Article Snippet: Slides were cover slipped with mounting medium containing 4,6-diamidino-2-phenylindole (DAPI; Vector Laboratories Inc., Burlingame, CA, USA) to label nuclei.

Techniques: Translocation Assay, Immunocytochemistry, Staining, Fluorescence, Western Blot

Journal: The World Journal of Men's Health

Article Title: Vactosertib, a Novel, Orally Bioavailable Activin Receptor-Like Kinase 5 Inhibitor, Promotes Regression of Fibrotic Plaques in a Rat Model of Peyronie's Disease

doi: 10.5534/wjmh.190071

Figure Lengend Snippet: Vactosertib decreases transforming growth factor-β (TGF-β1)-induced production of hydroxyproline and extracellular matrix proteins. Fibroblasts were pretreated for 24 hours with Vactosertib (400 ng/mL) then exposed with TGF-β1 (10 ng/mL) for 48 hours. (A) Hydroxyproline assay. Each bar depicts the mean value (±standard errors) of four experiments per group. a p<0.05, compared with the control group; b p<0.05, compared with the TGF-β1 group. (B) Representative Western blot for plasminogen activator inhibitor-1 (PAI-1), collagen I, collagen III, collagen IV, and fibronectin expressions in fibroblasts. (C) Data are presented as the relative density of each protein normalized to β-actin. Each bar depicts the mean value (±standard errors) of four experiments per group. a p<0.05, compared with the control group; b p<0.05, compared with the TGF-β1 group. (D) Representative fluorescent immunocytochemistry of fibroblasts with antibodies against PAI-1, fibronectin, collagen I, and collagen IV. Nuclei were stained with DAPI (4,6-diamidino-2-phenylindole). Scale bar indicates 100 µm. The results were similar among four independent experiments.

Article Snippet: Slides were cover slipped with mounting medium containing 4,6-diamidino-2-phenylindole (DAPI; Vector Laboratories Inc., Burlingame, CA, USA) to label nuclei.

Techniques: Hydroxyproline Assay, Western Blot, Immunocytochemistry, Staining