Figure 5.

HS and CS chains are responsible for endocytosis of 3D8 scFv. At 24 h post-transfection with plasmids encoding SDC2-GFP, HA-GPC3, CD44-GFP, or HA-BCAN, HeLa cells were treated with 500 mIU/ml heparinase III or 500 mIU/ml chondroitinase ABC for 2 h at 37 °C. Then, cells were treated for 6 h at 37 °C with 3D8 scFv, followed by immunoprecipitation with anti-GFP, anti-GPC3, anti-CD44, or anti-HA antibodies. Proteins in the extract (Input; 10%) and pulled-down fractions (IP) were analyzed by immunoblotting.