Figure 4: Cell-based assay of BoNT/C1 ad cleavage of SNAP-25 or Syx1.
From: Engineering Botulinum Neurotoxin C1 as a Molecular Vehicle for Intra-Neuronal Drug Delivery
Primary cortical neurons maintained for 14 days after plating were exposed to BoNT/C1 ad, wt BoNT/C1, or medium alone for 96 hours, and analyzed by Western blotting. lane 1 – neurons were treated with BoNT/C1 ad (5 nM, equivalent to 0.01 mouse LD50 units/mL); lane 2 – neurons were treated with BoNT/C1 ad (25 nM, equivalent to 0.05 mouse LD50 units/mL), lane 3 – neurons were treated with BoNT/C1 ad (100 nM, equivalent to 0.2 mouse LD50 units/mL), lane 4 – neurons were treated with wt BoNT/C1 (0.5 nM, equivalent to 405 mouse LD50 units/mL) (positive control), lane 5 – neurons were treated with medium alone (negative control). Immunoblots were probed with mAbs against SNAP-25 and Syx1. Immunostaining with mAbs against VAMP-2 and β-actin were used as loading controls. Western blots have been cropped to only show bands visualized in each independent experiment.
