Figure 6.

Assessment of the mechanism of CTX-Fc-BNCs internalization. A172 cells were treated with CTX-Fc-BNCs or human IgG-BNCs in the presence of 100 nM CPZ or 5 mM MβCD at 37°C for 1 h, followed by trypsinization. The cytoplasmic fraction was immunoprecipitated with anti-HBsAg antibody conjugated to microbeads. “Untreated” indicates that the cells were treated with CTX-Fc-BNCs in the absence of any inhibitors. (a) The precipitates were immunoblotted and detected using anti-human-IgG-Fc antibody. (b) The BNC bands were analyzed densitometrically using a CS Analyzer 3.0 and plotted in each graph to evaluate the amount endocytosed. U: untreated; CP: CPZ; Mβ: MβCD. The data (mean ± S.D) presented are from three independent experiments (**P < 0.05).