Figure 3

DDV entry into neurons via BoNT/A receptor mediated endocytosis. Images in rows A and B were obtained from triplicate cultures exposed for 16 h under the following conditions: (A1) DDV (200 nM) in the absence of rHC; (A2) DDV (200 nM) and rHC (200 nM); (A3) DDV (200 nM) and rHC (600 nM); and (A4) DDV (200 nM) and rHC (2 μM); (B1) DDV (200 nM) in the absence of BoNT/A; (B2) DDV (200 nM) and BoNT/A (200 nM); (B3) DDV (200 nM) and BoNT/A (600 nM); and (B4) DDV (200 nM) and BoNT/A (2 μM). DDV and rHC or BoNT/A were added simultaneously. Note the progressive reductions in fluorescence with increasing concentrations of BoNT/A or rHC. Micrographs were obtained on a Bio-Rad 2000 laser microscope confocal microscope using a 100× oil immersion objective. The fluorescence colors of labelled molecules are: red-rHC; green-OG488-dextran.