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. Author manuscript; available in PMC: 2011 Feb 1.
Published in final edited form as: Toxicon. 2009 Oct 21;55(2-3):619. doi: 10.1016/j.toxicon.2009.10.019

Fig. 4.

Fig. 4

Defined cationic polymer DNA transfection reagents promote BoNT intoxication independent of DNA transfection. A. N2A cells were incubated for 24 hrs with 20 nM recombinant BoNT/A LC (Lc438) and 0.5 µg pcDNA/CFP expression plasmid with (+) or without (−) pre-incubation with FuGene-HD (FuGene) or PEI having different average molecular weights as indicated. Pre-incubations were performed at a toxin (µg): reagent (µl) ratio of 1:1 with PEI and 1:3 with FuGEne HD. B. N2A cells were incubated for 24 hrs with 20 nM recombinant BoNT/A Lc438 (10 nM for ratios 1:3 and 1:4) and 0.25µg pcDNA/CFP following pre-incubation with FuGene-HD (FuGene) or PEI. (25000 or 75000 MW) at the indicated toxin (µg): reagent (µl) ratio. Cell extracts were prepared and the extent of BoNT/A intoxication was assessed by Western blotting to monitor SNAP25 cleavage. The efficiency of DNA transfection was assayed by Western blotting for CFP.