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Figure 5


Fig. 5. Co-distribution analysis of STxB and CTxB. (A,B) Immunofluorescence analysis of STxB (red) and CTxB (green) distribution in (A) Syn16 RNAi or (B) Syn5 RNAi cells after internalization for 45 minutes at 37°C. (C) Competition analysis. Iodinated STxB was incubated with HeLa cells in the presence of increasing concentrations of non-labeled CTxB or STxB. The amount of cell-associated radioactivity under each condition was determined. Note that CTxB did not displace the iodinated STxB from its receptor. (D) Binding analysis. HeLa cells were incubated on ice with 1 µM Alexa Fluor-488-labeled CTxB and 1 µM Cy3-labeled STxB. The cells were washed and fixed, and numbers of cells positive for one or the other marker were counted. Double-positive cells are roughly present at numbers as expected if GM1 and Gb3 expressions are independent events.





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