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. 2016 Jul 18;214(2):167–179. doi: 10.1083/jcb.201511102

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© 2016 Tang et al.

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Figure 8. — Modified ensemble motility assay showing that Myo51–Rng8/9 slides actin-Tpm filaments relative to each other. (A) Schematic of Myo51 attached to surface-immobilized green actin-Tpm (labeled with BODIPY-phallacidin) via the Rng8/9 appendage. The green actin-Tpm is bundled as Myo51–Rng8/9 cross-links filaments. Some Myo51 motor domains remain available to move red actin-Tpm filaments (labeled with rhodamine-phalloidin), whereas others are involved in bundle formation. (B and C) Top: kymographs of a red actin-Tpm filament that traveled along the green actin bundles formed by Myo51–Rng8/9. The white arrowheads indicate the moving red actin-Tpm filament at various time points. Note that in B, the filament paused midway in the trajectory and resumed motility. Bottom: maximum projection of the 60 s motility shown in the top panel. Bars, 5 µm. The red filament trajectory overlaps with the ridge of green actin-Tpm bundles. (D) Motile red filament speed distribution (n = 114). The mean ± SD is indicated.