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Sequential recruitment of EGFP-Aux1 and TagRFP-GAK to clathrin-coated vesicles. Bottom surface of a SUM159 cell gene-edited for EGFP-Aux1+/+ and TagRFP-GAK+/+ was imaged by TIRF microscopy every 1 s for 120 s. To facilitate visualization, the TagRFP channel was shifted laterally by 5 pixels in the right panel.
