Fig. 3

Raft localization of CD44 after TGF-β1 stimulation was inhibited by reduced sialylation with BGN. a, b Western blot analysis of phosphorylated extracellular signal-related kinase (pERK) was performed in control (Ctr; vehicle-treated EP fibroblasts in DMSO) and GalNAc-α-O-benzyl (BGN; 2 mM)-treated EP fibroblasts. The histogram (b) shows the mean densitometric analysis ± SD for the phosphorylated proteins normalized to the loading control. Values were obtained from three independent experiments. **P < 0.02. c Immunocytochemical staining was performed in control and BGN (2 mM)-treated EP fibroblasts after transforming growth factor (TGF)-β1 treatment. Representative images are shown (GM1, red; CD44, green; DAPI, blue; GM1 and CD44 colocalization, yellow). d The histogram shows the mean ± SD percentage of GM1-CD44 colocalized cells colored yellow, as shown in (c), from two independent experiments (total of six fields); ***P <0.01. e Immunoprecipitation (IP) of CD44 followed by immunoblotting of epidermal growth factor receptor (EGFR) was performed. Representative images are shown. f Total cell lysates from control and BGN-treated EP fibroblasts were pulled down with ECA. Immunoblotting of CD44 was performed on ECA-binding proteins. In addition, IP of CD44 followed by immunoblotting of ECA, MAL-II, and CD44 was performed. Representative images are shown