Role of actin polymerization and actin cables in actin-patch movement in Schizosaccharomyces pombe

Movie 1 (mov 397)
Three-dimensional images of the S. pombe actin cytoskeleton.
Rotating image of a three-dimensional reconstruction of wild-type cells stained with phalloidin-conjugated AlexaFluor 488 to visualize the actin cytoskeleton. Cells are slightly flattened because of pressure from the coverslip. Individual frames are pre-sented sequentially at projections of 2° angles, rotating about the y-axis.

Movie 2 (mov 404)
Three-dimensional images of the S. pombe actin cytoskeleton.
Rotating image of a three-dimensional reconstruction of wild-type cells as in Movie 1, rotating about the x-axis.

Movie 3 (mov 177)
Actin patches are highly dynamic.
Time-lapse recording of actin-patch dynamics in S. pombe cells expressing Crn1p−GFP, which labels actin patches. Images are two-dimensional projections of 5 optical sections (1 m). The time inter-val between frames is 5 s. Scale bar represents 5 m.

Movie 4 (mov 189)
Actin patches are located both in the cytoplasm and in association with the cell cortex. Series of optical sections of a wild-type cell expressing Crn1p−GFP. Eighteen optical sections are spaced 0.2 m apart and go from the upper cell surface to the lower. Scale bar represents 5 m.

Movie 5 (mov 165)
Actin patches exhibit directed and non-directed movements.
Time-lapse recording of actin patches exhibiting directed movement along linear tracks, and non-directed movement at cell tips. The time interval between frames is 0.5 s. The first frame is represented by Fig. 2a, 0 s. Scale bar represents 5 m.

Movie 6 (mov 246)
Time-lapse recording of an actin patch exhibiting directed movement along a Crn1p−GFP cable.
The time interval between frames is 0.5 s. The first frame is rep-resented by Fig. 2a, 0 s. Scale bar represents 5 m.

Movie 7 (mov 770)
Movement of actin patches requires actin polymerization.
Time-lapse recording showing inhibition of actin-patch movement after attenuation of actin polymerization. Cells were treated with 50 m LatA for 2 min and movement of Crn1p−GFP patches in live cells was then recorded. Inhibition of actin polymerization with LatA abolishes actin-patch movement. The time interval between frames is 0.5 s. The first frame is represented by Fig. 3a, left panel. Scale bar represents 5m.

Movie 8 (mov 199)
Wild-type control treated with 1% dimethylsulphoxide (DMSO).
Control time-lapse recording of actin patch-movement after treatment with 1% DMSO for 2 min. This treatment does not affect the pattern or rate of actin-patch movement. The time interval between frames is 0.5 s. The first frame is represent-ed by Fig. 3a, right panel. Scale bar represents 5 m.

Movie 9 (mov 743)
Arp3 is required for movement of actin patches.
Time-lapse recording showing reduction of actin-patch movement in the cold-sensitive arp3 mutant. Cells were shifted to 19 °C for 100 min before imaging Crn1p−GFP fluorescence. The time interval between frames is 0.5 s. The first frame is represented by Fig. 4a, arp3. Scale bar represents 5 µm.

Movie 10 (mov 210)
Profilin is required for movement of actin patches.
Time-lapse recording showing reduction of actin-patch movement in the temperature-sensitive cdc3 (profilin) mutant. Cells were shifted to 36 °C for 4 h before imaging Crn1p−GFP fluorescence. The time interval between frames is 0.5 s. The first frame is represented by Fig. 4a, cdc3. Scale bar represents 5 m.

Movie 11 (mov 1,442)
Microtubules are not required for movement of actin patches.
Time-lapse recording showing that actin-patch movement is unaffected in cells treated with 25 mg ml−1 MBC for 10 min to depolymerize microtubules. The time interval between frames is 0.5 s. The first frame is represented by Fig. 4a, +MBC. Scale bar represents 5 m.

Movie 12 (mov 1,854)
Wild-type control treated with 1% DMSO.
Control time-lapse record-ing of actin-patch movement after treatment with 1% DMSO for 10 min. This treat-ment does not effect the pattern or rate of actin-patch movement. The time interval between frames is 0.5 s. The first frame is represented by Fig. 4a, + 1% DMSO. Scale bar represents 5 m.

Movie 13 (mov 1,898)
Wild-type 36 °C control.
Control time-lapse recording of actin-patch movement after a shift to 36 °C for 4 h. Incubation at 36 °C does not effect the pattern or rate of actin-patch movement. The time interval between frames is 0.5 s. The first frame is represented by Fig. 4a, 36 °C. Scale bar represents 5 m.

Movie 14 (mov 1,757)
Wild-type 19 °C control.
Control time-lapse recording of actin-patch movement after a shift to 19 °C for 100 min. Incubation at 19 °C does not effect the pattern or rate of actin-patch movement. The time interval between frames is 0.5 s. The first frame is represented by Fig. 4a, 19 °C. Scale bar represents 5 m.

Movie 15 (mov 252)
Directional movement of actin patches requires tropomyosin.
Time-lapse recording showing loss of directed actin-patch movement in the temperature- sensitive cdc8 (tropomyosin) mutant, which lacks actin cables. Cells were shifted to 35.5 °C for 25 min before imaging Crn1p−GFP fluorescence. The time interval between frames is 0.5 s. The first frame is represented by Fig. 5a, cdc8. Scale bar represents 5 m.

Movie 16 (mov 473)
Wild-type 35.5 °C control after 25 min.
Control time-lapse recording of actin-patch movement after a shift to 35.5 °C for 25 min. Incubation at 35.5 °C does not effect the pattern or rate of actin-patch movement. The time interval between frames is 0.5 s. The first frame is represented by Fig. 5a, wild type. Scale bar represents 5 m.

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