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Live-cell imaging was performed using a Nikon Eclipse Ti-E inverted microscope equipped with a TIRF 60x objective (Micro Video Inst., Avon, MA), a Cascade 512b EMCCD camera (Photometrics, Tucson, AZ), and a chameleon-2 filter set (Chroma Technology Corp., Bellows Falls, VT) comprised of a D440/20 excitation filter, a 455DCLP dichroic, and D485/40 (CFP) or D535/30 (YFP) emission filters 34,35