Fig. 5

Defect in raft localization of CD44 during the process to senescence was restored by treatment with a sialidase inhibitor. a Cell surface glycans in early passage (EP), late passage (LP), and zanamivir (2 mM)-treated LP fibroblasts were analyzed by FACS using lectins. Mean fluorescent intensities (MFIs) relative to those of control cells are shown. Results are presented as means ± SD from three independent experiments. b Total cell lysates from EP, LP, and zanamivir-treated LP fibroblasts were pulled down with ECA. Immunoblotting of CD44 was performed on ECA-binding proteins (left). In addition, immunoprecipitation (IP) of CD44, followed by immunoblotting of ECA, MAL-II, and CD44, was performed (right). Representative images are shown. c Immunocytochemical staining was performed in EP, LP, and zanamivir-treated LP fibroblasts after transforming growth factor (TGF)-β1 treatment. Representative images are shown (GM1, red; CD44, green; DAPI, blue; 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 IP of CD44 followed by immunoblotting of epidermal growth factor receptor (EGFR) was performed. Representative images are shown