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Fig. 3. Integrins are concentrated in nascent adhesions demarked by the presence of paxillin-containing adhesions. MEF or CHO cells were transfected with both paxillin-CFP (A) and {alpha}5-YFP (B). Confocal image time series were collected 24 hours after transfection and 1 hour after cells were plated on coverslips coated with 1 µg ml–1 fibronectin. The % increase in the intensity of {alpha}5-YFP in nascent adhesive areas relative to adjacent areas was calculated and plotted as a histogram (C; 144 adhesions, 17 cells). In 5% of the adhesions measured, {alpha}5-integrin showed an equal or slightly lower intensity of fluorescence in areas adjacent to paxillin containing adhesions. (D) The relative concentration of {alpha}5-integrin in adhesive areas is higher for lower-expressing cells. For the two image frames shown there is a 27% increase in {alpha}5 intensity under paxillin based on measurements taken on 20 adhesions. Scale bar, 5 µm.