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Fig. 3. Colocalization of dynamin2-EGFP in dsRed-clathrin puncta in migrating MDCK cells. (A) dsRed-clathrin epifluorescence image. (B) Dynamin2-EGFP epifluorescence image. (C) Overlay of A and B. Scale bar, 5 µm. (D) dsRed-clathrin TIR-FM image. (E) Dynamin2-EGFP TIR-FM image. (F) Overlay of D and E. Scale bars, 5 µm. (G) Depiction of the decrease of mean ± s.e.m. of correlation coefficient comparing images in D and E following pixel shift in all four directions. (H) Graph of the dynamin2-EGFP fluorescence relative to dsRed-clathrin fluorescence (normalized to whole cell relative fluorophore intensity for each cell) within 150 clathrin puncta from three cells (50 per cell).