Fig. 5. Dependency of auxilin and Hsc70 concentration on the uncoating of clathrin-coated pits. (A) Different concentrations of auxilin (between 2x10⁵ µM and 2 µM) were added to permeabilized cells in the presence of 1 mM ATP and 2 µM Hsc70. (B) Different concentrations of Hsc70 (between 2x10⁴ µM and 10 µM) were added to permeabilized cells in the presence of 1 mM ATP and 0.2 µM auxilin. Cells were imaged at 30-second intervals for 5 minutes. Cells of similar fluorescence intensity were analyzed. The decrease in relative fluorescence intensity was measured as a function of time. Data were normalized to the maximal initial fluorescence in each plot.