Figure 2

Expression of ERM proteins in TG cells, and the binding capacity between ezrin and Hsc70. (A) Expression of ERM proteins in TG, TS, and J774 macrophage cells. Immunoblot analyses were performed with anti-ezrin, anti-radixin, and anti-moesin antibodies. (B) Affinity of ERM proteins for Hsc70 shown by pull-down assay. Recombinant Hsc70 and each ERM protein were mixed, and these samples were immunoprecipitated with anti-Hsc70 antibodies or beads only. Detection of proteins was performed by immunoblotting. (C) The binding capacity of ERM proteins to Hsc70 were measured by ELISA. Immunoplates were coated with each ERM protein or BSA (control), and then Hsc70 was added. Data are the averages of triplicate samples from three identical experiments. Error bars represent standard deviation. Statistically significant differences between control and ERM proteins are indicated by asterisks (*, P < 0.01). (D) ELISA was used to determine the binding domain of Hsc70 for ezrin. Immunoplates were coated with full-length Hsc70 (Full Hsc70), ATP-binding domain (ATPase), and peptide-binding domain (PBD) of Hsc70, following which ezrin was added. Data are the averages of triplicate samples from three identical experiments. Error bars represent standard deviation. Statistically significant differences between control and each domain proteins are indicated by asterisks (*, P < 0.01).