Imaging was performed on a Nikon Ti Eclipse microscope quipped with a TIRF module coupled to Agilent monolithic laser combiner MLC400 (laser lines: 405, 488, 561, and 653 nm; Agilent Technologies). Images were collected with a 100 × 1.49 numerical aperture (NA) objective with an electron multiplying charge-coupled device camera (Evolve 512, Photometrics), yielding a pixel size of 103 nm. Multicolor, time lapse, and stream acquisition were controlled and recorded using the software μManager. Image analysis was performed in ImageJ (

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