FIGURE 2.

Generation of GFP-VAMP2 SiMa cell line using viral transduction. (A) Schematic showing the fusion protein containing GFP inserted at the amino terminus of VAMP2. TMR, transmembrane region. (B) GFP-VAMP2 localizes to vesicular structures (top) and is released into the cytosol (bottom) upon incubation with BoNT/B (30 nM). (C) Robust expression following viral transduction and puromycin selection is evident in GFP and differential interference contrast (DIC) merged images. (D) Stable GFP-VAMP2 cleavage product can be detected following incubation with BoNT/B (30 nM). Immunoblotting was performed using a GFP antibody.