JCS03048 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Cytology of cells under investigation and authentication of GFP-AtEB1 labelling to microtubule plus ends. (A) Relative cell numbers of wild-type, GFP-AtEB1 and GFP-TUA6 transformed BY-2 cells. (B-F) GFP-AtEB1 (green) labelling to the microtubule plus ends of YFP-MAP4 (red) labelled microtubules, which are growing (yellow arrowheads) and apparent loss of GFP-AtEB1 from microtubule plus ends upon initiation of microtubule shrinking (yellow arrows). Bar, 1 μm. Time is indicated in seconds.

• Supplemental Figure 2 -

  Fig. S2. Whole-spindle kymograph analysis of areas near chromosomes and areas with overlapping microtubule but without chromosomes. The spindle micrograph shown in Fig. 3B was subdivided into different x positions numbered 7 to 42 (A) from which kymographs (B) were created. Rectangles adjacent to (negatively stained) chromosomes are red, others are blue. Rectangles of the same colour were mounted into a single image, which subsequently was transformed to Fourier space. Green dots indicate that rectangles were flipped vertically before being mounted. The Fourier transformation representing all red rectangles is shown in Fig. 3E. The Fourier transformation representing all blue rectangles is shown in Fig. 3K.

• Supplemental Figure 3 -

  Fig. S3. Amount of microtubules close from the equator and at poles. (A,B) GFP-MAP4-labelled metaphase spindle microtubules. Red rectangle indicates the area from which the GFP-MAP4 intensity profile (shown in B) is obtained. Bar, 5 μm.

• Movie 1 -

  Movie 1. Dynamics of GFP-AtEB1-labelled microtubule plus ends during spindle formation. The movie represents xyt-scanning containing 406 frames acquired at a scanning speed of 2 seconds/frame (see also Fig. 2A-J).

• Movie 2 -

  Movie 2. Dynamics of GFP-AtEB1-labelled microtubule plus ends in metaphase plant spindle. The movie represents xyt-scanning containing 134 frames acquired at a scanning speed of 2 seconds/frame (see also Fig. 3 and Fig. 6).

• Movie 3 -

  Movie 3. Dynamics of GFP-AtEB1-labelled microtubule plus ends and SYTO-labelled chromosomes at the onset of anaphase in plant cells. The movie represents xyt-scanning containing 116 frames acquired at a scanning speed of 5.5 seconds/frame (see also Fig. 7).

• Movie 4 -

  Movie 4. Translocation of photobleached areas in case of line photobleaching performed equidistant from the spindle equator during metaphase in cells expressing GFP-α tubulin. The movie represents xyt-scanning containing 56 frames acquired at a scanning speed of 2 seconds/frame (see also Fig. 8A-D).

• Movie 5 -

  Movie 5. Translocation of photobleached areas in case of line photobleaching performed equidistant from the spindle equator during anaphase in cells expressing GFP-α tubulin. The movie represents xyt-scanning containing 54 frames acquired at a scanning speed of 2 seconds/frame (see also Fig. 8E-H).