Fig. 4

Myofibroblast differentiation was inhibited by sialidase. a, b Cell surface glycans in control (Ctr; non-treated EP fibroblasts) and 100 U/ml sialidase-treated EP fibroblasts were analyzed by FACS using lectins. Three independent experiments were performed and representative results are shown (a). Controls are presented in gray. Mean fluorescent intensities (MFIs) relative to those of control cells are shown (b). Results are presented as means ± SD from three independent experiments. c, d Western blot analysis of α-smooth muscle actin (α-SMA) was performed 3 days after myofibroblast differentiation in control and sialidase-treated EP fibroblasts. The histogram (d) shows the mean densitometric analysis ± SD of α-SMA normalized to the loading control (β-actin). The values were obtained from three independent experiments. e Immunocytochemical staining was performed in control and sialidase-treated EP fibroblasts after transforming growth factor (TGF)-β1 treatment. Representative images are shown (GM1, red; CD44, green; DAPI, blue; colocalization, yellow). f The histogram shows the mean ± SD percentage of GM1-CD44 colocalized cells colored yellow, as shown in (e), from two independent experiments (total of six fields); ***P <0.01. g Immunoprecipitation (IP) of CD44 followed by immunoblotting of epidermal growth factor receptor (EGFR) was performed. Representative images are shown