Figure 3. Lung myofibroblasts demonstrate increased α6-expression.

(a) Frozen lung tissue sections obtained from failed normal human donors, patients with IPF, saline-treated mice and bleomycin-treated mice were double-stained for α₆ (green) and αSMA (red). Nuclei were stained by DAPI (blue). Confocal immunofluorescent images were overlaid to show α₆-expression in αSMA-positive lung myofibroblasts. Scale bar, 50 μm; scale bar, 20 μm for mouse with bleo images. (b) Comparison for α₆-expression in lung (myo)fibroblasts isolated from patients with IPF (n=10) and non-ILD control human subjects (n=6) by immunoblot; Relative levels of α₆-protein normalized to GAPDH expression. Results are the means±s.d. Representative blots for α₆-expression as well as β₁- and β₄-expression were shown. A549 cells were used as positive control for β₄-expression in immunoblot analysis. Relative levels of α₆-, β₁- and β₄-expression on the cell surface of IPF lung myofibroblasts were analysed by flow cytometry. (c) Detection of α₆β₁- and α₆β₄-complexes in IPF lung myofibroblasts by immunoprecipitation and immunoblot. (d) Identification of α₆A and α₆B expression in human and mouse lung tissues and fibroblasts by immunoblot; *P<0.05, one-way analysis of variance.