Chlorotoxin Fused to IgG-Fc Inhibits Glioblastoma Cell Motility via Receptor-Mediated Endocytosis
Figure 5
Evaluation of CTX-Fc-BNCs internalized by A172 cells. (a) A172 cells were incubated with CTX-Fc-BNCs at 37°C. In “CTX competition”, the cells were treated primarily with CTX at 4°C for 20 min before incubating with CTX-Fc-BNCs. The cells were stained with anti-human IgG antibody labeled with FITC. Left: fluorescence image; right; composite image. Bars = 10 μm. (b) and (c) A172 cells were treated with CTX-Fc-BNCs, human IgG-BNCs, or M-CTX-Fc for 1 h at 4°C or 37°C. After incubation, the cells were trypsinized. The cytoplasmic fraction was immunoprecipitated with anti-HBsAg antibody conjugated to microbeads. (b) The precipitates were immunoblotted and detected with anti-human-IgG-Fc antibody. (c) The BNC bands in the CTX-Fc-BNCs or human IgG-BNCs treatment at 37°C were analyzed densitometrically using a CS Analyzer 3.0 and plotted in each graph to estimate the amount endocytosed. The data (mean S.D) presented are from three independent experiments ().
