Supplemental Movie Legends
Supplemental Movie 1. Reorganization of Cp-Actin Filaments during the Blue Light-Induced Chloroplast Avoidance Response.
The same cell as in Figure 1A is shown. Time-lapse images (maximized with three images in a 1.2-µm depth) were collected at approximately 31-s intervals and played back at 5 frames per second (fps). The total elapsed time is 15:56 (mm:ss). The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. The region indicated with the blue rectangle (15 µm x 50 µm) was irradiated using 458-nm laser scans during the intervals between the image acquisitions to induce the avoidance response. Scale bar = 10 µm.
Supplemental Movie 2. Reorganization of THRUMIN1-GFP during the Blue Light-Induced Chloroplast Avoidance Response.
The same cell as in Figure 2B is shown. Time-lapse images (maximized with three images in a 1.2-µm depth) were collected at approximately 31-s intervals and played back at 5 fps. The total elapsed time is 08:16 (mm:ss). The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. The region indicated by the blue rectangle was irradiated using 458-nm laser scans during the intervals between the image acquisitions. Scale bar = 5 µm.
Supplemental Movie 3. Reorganization of Cp-Actin Filaments in athrumin1 Mutant Cell under a Condition Inducing the Blue Light-Induced Chloroplast Avoidance Response.
The same cell as in Figure 2C is shown. Time-lapse images (maximized with three images in a 1.2-µm depth) were collected at approximately 31-s intervals and played back at 5 fps. The total elapsed time is 15:30 (mm:ss). The region indicated by the blue rectangle was irradiated using 458-nm laser scans during the intervals between the image acquisitions. The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. The paths of the chloroplasts were traced and are shown in colored circles. Scale bar = 5 µm.
Supplemental Movie 4. Reorganization of THRUMIN1-GFP in a chup1 Mutant Cell under a Condition Inducing the Blue Light-Induced Chloroplast Avoidance Response.
The same cell as in Figure 2D is shown. Other details are the same as for Supplemental Movie 3.
Supplemental Movie 5. Reorganizations of Cp-Actin Filaments on the Leading Edge of Chloroplasts during an Avoidance Response.
The same cell as in Figure 3 is shown. Time-lapse images (maximized with three images in a 1.5-µm depth) were collected at approximately 34.5-s intervals and played back at 5 fps. The total elapsed time is 12:04 (mm:ss). The region indicated by the blue circle(20 µm in diameter) was irradiated using 458-nm laser scans during the intervals between the image acquisitions. The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. The yellow rectangular box denotes the region of the images shown in Figure 3E. Scale bar = 10 µm.
Supplemental Movie 6. Reorganization of Cp-Actin Filaments during the Accumulation Response Following the Avoidance Response.
The same cell as in Figure 4A is shown. Time-lapse images (maximized with three images in a 1.5-µm depth) were collected at approximately 36.5-s intervals and played back at 5 fps. The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. The region indicated by the blue rectangle was irradiated using 458-nm laser scans during the intervals between the image acquisitions for the avoidance response, and the light was switched off for the accumulation response. Scale bar = 10 µm.
Supplemental Movie 7. Reorganizations of Cp-Actin Filaments and the Chloroplast Envelope during the Avoidance Response in Side-Viewed Chloroplasts.
The same cells as in Figure 5A (left, cp-actin probed with GFP-TALIN) and 5B (right, chloroplast envelope probed with OEP7-YFP) are shown. Time-lapse images (maximized with three images in a 1.2-µm depth) were collected at approximately 30-s (left) and 29-s (right) intervals, respectively and played back at 5 fps. The total elapsed time is 15:27 (left) and 14:57 (right) (mm:ss). The regions indicated by the blue rectangles(15 µm x 20 µm) were irradiated using 458-nm laser scans during the intervals between the image acquisitions. The images are false-colored to indicate GFP (green), YFP (yellow), and chlorophyll (red) fluorescence. Scale bars = 5 µm.
Supplemental Movie 8. Reorganization of Chloroplast Envelope proved by OEP7-YFP during the Avoidance Response.
The same cells as in Figure 5D are shown. Time-lapse images (maximized with three images in a 1.2-µm depth) were collected at approximately 31-s intervals and played back at 5 fps. The total elapsed time is 15:45 (mm:ss). The region indicated by the blue rectangle was irradiated using 458-nm laser scans during the intervals between the image acquisitions. The path of the chloroplast was traced and is shown in red circles. The images are false-colored to indicate YFP (yellow) and chlorophyll (red) fluorescence. Scale bar = 5 µm.
Supplemental Movie 9. Disappearance of the Cp-Actin Filaments Captured by CLSM in a WT Cell.
Time-lapse images were collected at approximately 0.95-s intervals using continuous scanning with 458-nm and 488-nm lasers and played back at 20 fps. The total elapsed time is approximately 03:00 (mm:ss). The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. Scale bar = 10 µm.
Supplemental Movie 10. Cp-Actin Filament Dynamics Captured by TIRFM in a WT Cell.
The same WT cell as in Figure 7 is shown. Time-lapse images were collected at approximately 0.2-s intervals using 488-nm laser excitation and played back at 50 fps. The total elapsed time is 02:59 (mm:ss).
Supplemental Movie 11. Cp-Actin Filament Dynamics Captured by TIRFM in a phot1 Mutant Cell.
The samephot1 cell as in Figure 7 is shown. Other details are as in Supplemental Movie 10.
Supplemental Movie 12. Cp-Actin Filament Dynamics Captured by TIRFM in a phot2 Mutant Cell.
The same phot2 cell as in Figure 7 is shown. Other details are as in Supplemental Movie 10.
Supplemental Movie 13. Cp-Actin Filament Dynamics Captured by TIRFM in a phot1 phot2 Mutant Cell.
The same phot1 phot2 cell as in Figure 7 is shown. Other details are as in Supplemental Movie 10.
Supplemental Movie 14. Cp-Actin Filament Dynamics Captured by TIRFM in a chup1 Mutant Cell.
The same chup1 cell as in Figure 7 is shown. Other details are as in Supplemental Movie 10.
Supplemental Movie 15. Reorganization of the Cp-Actin Filaments in Mock-Treated Cells for 1 h.
The same cellasin Figure 9A 0 mM i-iv is shown. Time-lapse images (maximized with three images in a 1.2-µm depth) were collected at approximately 31-s intervals using 488-nm laser excitation and played back at 5 fps. The total elapsed time is approximately 15:40 (mm:ss). The region (10 µm x 25 µm)indicated by the blue rectanglewas irradiated using 458-nm laser scans during the intervals between the image acquisitions to induce the avoidance response. The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. Scale bar = 25 µm.
Supplemental Movie 16. Reorganizations of the Cp-Actin Filaments in 50 mM BDM-Treated Cells for 1 h.
The same cell as inFigure 9A 50mM (1h) v-viii is shown. Other details are as in Supplemental Movie 15.
Supplemental Movie 17. Cp-Actin Filament Dynamics Captured by CLSM in a Cell Treated with 50 mM BDM for 1 h.
The same cell in as Figure 9A and Supplemental Movie 16 is shown. Time-lapse images were collected at approximately 1.05-s intervals using continuous scanning with 458-nm and 488-nm lasers after microbeam irradiation with a 458-nm laser (Supplemental Movie 16) and played back at 20 fps. The total elapsed time is approximately 03:00 (mm:ss). The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. Scale bar = 25 µm.
Supplemental Movie 18. Reorganizations of the Cp-Actin Filaments in 50 mM BDM-Treated Cells for 2 h.
The same cell as in Figure 9A 50mM (2h) ix-xii is shown. Other details are as in Supplemental Movie 15.
Supplemental Movie 19. Cp-Actin Filament Dynamics Captured by CLSM in a Cell Treated with 50 mM BDM for 2 h.
The same cell as in Figure 9A and Supplemental Movie 18 is shown. Other details are as in Supplemental Movie 17.
Supplemental Movie 20.Disappearance of the Fragmented Cp-Actin Filaments via the Chloroplast Edge.
Time-lapse sequence from 20 s (101th frame) to 60 s (300th frame) as shown in Supplemental Movie 11. The time-lapse images were played back at 30 fps. The total elapsed time is 40 s. The images are false-colored to indicate GFP (green) and chlorophyll (red) fluorescence. Note that most of the fragmented cp-actin filaments moved to the chloroplast edge and disappeared (see selected tracks of representative cp-actin fragments).