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. Author manuscript; available in PMC: 2011 Nov 1.
Published in final edited form as: Toxicon. 2010 Jul 14;56(6):990–998. doi: 10.1016/j.toxicon.2010.07.003

Fig. 6.

Fig. 6

VHH ALc-B8 inhibits BoNT/A protease when co-expressed within cultured neuronal cells. (A) M17 cells were co-transfected with an expression vector encoding the indicator protein, YFP/SNAP25/CFP, and either a control plasmid (−) or an expression plasmid for YFP/ALc-B8 (+) as indicated. After 24 h post-transfection, the cells were exposed to 10 nM BoNT/A (+) or media (−). Cell extracts were prepared 24 h later and the cleavage of indicator by BoNT/A was detected by Western blot with anti-GFP antibody which recognizes both the indicator and the YFP/ALc-B8. The weak band co-migrating with ALc-B8 in the control cells is a BoNT/A cleavage product of the indicator protein. (B) M17 cells were transfected with either a control plasmid (−) or an expression plasmid for YFP/ALc-B8 (+). 24 h later, transfected cells were exposed to 10 nM of BoNT/A (+) or media (−). Cell extracts were prepared 24 h later and BoNT/A cleavage of endogenous SNAP25 was detected by Western blot with anti-SNAP25 antibody (Sigma).