Transforming growth factor-β1 induces phosphorylation of Smad2 and epithelial–mesenchymal transition in BPH-1 cells. The cells were lysed using radioimmunoprecipitation assay buffer, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunoblotting detection of p-Smad2 (Ser465/467); Smad2 and vimentin was performed. Detection of β-actin served as control of equal loading. Densitometric measurements were performed using ImageJ software (NIH, Bethesda, MD, USA) and normalized to the expression of β-actin. The bar graph represents the average optical density.