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Figure 3


Fig. 3. Ligand binding to the {alpha}4 integrin extracellular domain regulates its association with paxillin. (A) B16F1 mouse melanoma cells were transfected with {alpha}4-integrin-GFP and paxillin-mRFP. Cells were then plated onto FN- or PLL-coated coverslips and imaged by multiphoton FLIM as before. (B) The same cells were plated onto H120-coated coverslips followed by treatment with DMF vehicle control or stated small molecule inhibitors for 30 minutes and imaged by multiphoton FLIM. Histogram analysis of relative spread of FRET efficiencies is a mean of 18 cells per treatment compared with {alpha}4-integrin-GFP alone control. (C) Human A375-SM melanoma cells were transfected with {alpha}4-integrin-GFP and paxillin-mRFP and plated onto a monolayer of TNF{alpha}-treated activated HUVEC cells. Cells were allowed to adhere for 60 minutes, and were then treated with DMF vehicle control or stated small molecule inhibitors for 30 minutes. Cells were then imaged using multiphoton FLIM as before.





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