Supplementary Materials

Actin Network Architecture Can Determine Myosin Motor Activity

Anne-Cécile Reymann, Rajaa Boujemaa-Paterski, Jean-Louis Martiel, Christophe Guérin, Wenxiang Cao, Harvey F. Chin, Enrique M. De La Cruz, Manuel Théry, Laurent Blanchoin

Materials/Methods, Supporting Text, Tables, Figures, and/or References

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  • Materials and Methods
  • Figs. S1 to S9
  • Full References

Images, Video, and Other Other Media

Movie S1
Actin filaments growing on a pWA-coated bar. Actin polymerization is followed by TIRF microscopy. (Left) Actin filaments are branched on the patterned region and parallel outside this region. (Right) A zoomed region (represented by the box in the left panel). Total elapsed time is 73 min, which is compressed to a 6-s QuickTime
Movie S2
Myosin VI–induced actin meshwork contraction and disassembly. Montage of time-series of myosin VI-induced network contraction on a bar-shaped micropattern. Total elapsed time is ~ 2 hours, which is compressed to a 13-s QuickTime movie. An example of such network contraction on a bar-shaped micropattern is illustrated in Fig. 1A.
Movie S3
Myosin II–induced actin meshwork contraction and disassembly. Montage of time-series of myosin II-induced network contraction on a bar-shaped micropattern. Total elapsed time is 158 min, which is compressed to a 27-s QuickTime movie. An example of such network contraction on a bar-shaped micropattern is illustrated in Fig. 1B.
Movie S4
Myosin VI–induced contraction and disassembly of an actin network nucleated from a ring-shaped micropattern and cross-linked with 100 nM α-actinin. Total elapsed time is 3 hours 33 min, which is compressed to a 7-s QuickTime movie.
Movie S5
Myosin VI–induced actin meshwork contraction and disassembly followed by TIRF microscopy. Same as movie S1 but in the presence of myosin VI in the reaction mixture. Total elapsed time is 618 min, which is compressed to a 6-s QuickTime movie, that corresponds to the time-lapse series in Fig. S4.
Movie S6
Tension and/or deformation of cohesive actin network by myosins VI and II. (Left) Actin filaments nucleated from a micropatterned bar (4-µm width), coated with pWA, elongate and undergo tension and deformation as they interact with HMM-myosin VI spotted on an adjacent micropattern. Total elapsed time is ~33 min, which is compressed to a 4-s QuickTime movie that corresponds to the time-lapse series in Fig. S5B. (Right) Actin filaments nucleated from a micropatterned bar (4-µm width), coated with pWA, elongate and undergo deformation as they interact with myosin II spotted on an adjacent micropattern. Total elapsed time is ~88 min, which is compressed to a 4-s QuickTime movie that corresponds to the time-lapse series in Fig. S5D.
Movie S7
(Left) Dynamic organization of actin architectures assembled on an eight-branch nucleating radial array in absence of myosin. Total elapsed time is ~1 hour 19 min, which is compressed to a 5-s QuickTime movie that corresponds to the time-lapse series in Fig. 2A. (Right) Regioselective myosin-induced contraction and disassembly of complex actin architecture. Kinetics of myosin-induced architecture selective contraction and disassembly, on an eight-branch actin nucleating radial array. As the denser entangled nucleating zones and the antiparallel links between them rapidly contract and disassemble, the parallel bundles keep on elongating without perturbation. Total elapsed time is ~1 hour 29 min, which is compressed to a 5-s QuickTime movie that corresponds to the time-lapse series in Fig. 2B.
Movie S8
Myosin-induced contraction of actin network nucleated on eight-branch nucleating radial arrays. Total elapsed time is 49 min, which is compressed to a 10-s QuickTime movie.
Movie S9
Myosin-induced contraction of actin network nucleated on full rings. The rings are 75 µm in diameter. Total elapsed time is 46 min, which is compressed to a 9-s QuickTime movie.
Movie S10
Comparison of myosin-induced contraction of actin network nucleated on a full (left) or dotted (right) ring. The rings are 75 µm in diameter. Total elapsed time is 19 min, which is compressed to a 10-s QuickTime movie that corresponds to the time-lapse series in Fig.3.
Movie S11
Scalability of ring contraction. Rings of 50 µm (left) and 75 µm (right) in diameter constituted of 12 nucleation dots (top) or 18 nucleation dots (bottom) were contracted by HMM–myosin VI at different velocities (see Fig. 4A for measurements). Total elapsed time is 20 min, which is compressed to an 11-s QuickTime movie.