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Figure 2


Fig. 2. Imaging of ligand pairs conjugated to different fluorophores. (A) COS-7 cells grown on coverslips were placed in KRH buffer at 35°C, and exposed to 10 µg/ml Alexa-Fluor-488-Tf + 10 µg/ml Alexa-Fluor-568-Tf (top panels), 50 ng/ml Alexa-Fluor-568-EGF + 50 ng/ml Alexa-Fluor-488-EGF (middle panels), or 50 ng/ml Alexa-Fluor-568-EGF + 10 µg/ml Alexa-Fluor-488-Tf (bottom panels) for 5 minutes. (Right panels) Merged images showing the overlap. (B) Images from the time periods after addition of ligands indicated above the panels. Arrows indicate the clear segregation of signals seen in cells incubated with different ligands, that is not seen with the same ligand conjugated to two different fluorophores. (C) Quantification of signal overlap between fluorophores after 5 minutes of exposure. Values are mean ± s.e.m. of five different experiments. Statistical significance of the differences between groups was estimated using two-tailed Student's t-tests.





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