Bleomycin-treated mice undergo robust epithelial to mesenchymal transition (EMT). (a) Representative photomicrographs of lung sections were obtained from mice treated with bleomycin (BLM) or saline (week 3). Sections were stained with hematoxylin and eosin (H&E), and Masson’s trichrome, as well as by immunohistochemistry for anti-α-smooth muscle actin(α-SMA), anti-collagen I (Col I), anti-collagen III (Col III), anti-transforming growth factor (TGF)-β antibody. μm (for H&E and Masson’s trichrome staining), or 50 μm (for immunohistochemistry). (b) Expressions of col1a2 and col3a1 from the lung tissues of BLM-treated mice were measured by real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis relative to that of saline-treated mice. (for col1a2); (for col3a1). (c) Protein levels of α-SMA, E-cadherin and GADPH in lung homogenates were determined by western blotting and analyzed by densitometry compared to GADPH expression. The same amounts of total protein are loaded in each lane. (d) Protein levels of phosphorylated Smad2 (pSmad2), total Smad2, and GADPH in lung homogenates were determined by western blotting and analyzed by densitometry compared to GADPH expression. The same amounts of total protein are loaded in each lane. Fold increase in pSmad2 expression after normalization to total Smad2 expression is shown. ; . Data presented in (b), (c), and (d) are representative of results from two independent experiments with five mice in each group. Values are shown as .
