Fig. 3. NTF-NgR generation is inhibited by BFA and occurs at the cell surface. (A) SH-SY5Y cells expressing NgR were incubated for 6 hours in OptiMEM in the absence (–) or presence (+) of 5 µM BFA. NgR was detected in the cell lysate and medium using the anti-V5 antibody. (B) Cells were surface biotinylated at 4°C and then incubated at 37°C in OptiMEM for the indicated times. Biotinylated proteins in the lysate and medium were precipitated with streptavidin-agarose and NgR detected with the anti-V5 antibody.