JCS02959 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Apoptotic microtubules are unmodified. Confocal image of anisomycin-treated HeLa cells labelled for tyrosinated α-tubulin (green) with acetylated α-tubulin (red). Bar, 10 μm.

• Supplemental Figure 2 -

  Fig. S2. Spike formation in UV-irradiated A431 cells. (A) Time-lapse microscopy of apoptosis in A431 cells showing apoptotic spike assembly during the late-execution phase. Frames are from supplementary material Movie 4. (B) Nocodazole prevents spike formation and cells undergo minimal fragmentation (frames from supplementary material Movie 5). (C) Spikes in the absence or presence of inhibitors. Bars, 10 μm.

• Supplemental Figure 3 -

  Fig. S3. Spikes are features of apoptosis in many cell-lines. (A) Formation of spikes in apoptotic HeLa cells. Apoptotic Jurkat (B), SW13-Cl.2 (C) and Meg01 cells (D) each showing numerous spikes. Bars, 2 μm (A); 10 μm (B,C,D).

• Supplemental Figure 4 -

  Fig. S4: Cartoon of microtubule organisation during the execution phase of apoptosis. Microtubules are depicted in red, and chromatin in blue.

• Movie 1 -

  Movie 1. Volocity 3-D reconstruction of microtubule (red) and chromatin (blue) organisation in an apoptotic HeLa cell.

• Movie 2 -

  Movie 2. Time-lapse movie of microtubule and chromatin dynamics in early apoptosis. A UV-treated A431 cell transiently-expressing YFP-tubulin (green) and HMGB1-CFP (red). Images were obtained at 300 second intervals. Movie plays at 2 frames/second.

• Movie 3 -

  Movie 3. GFP-Centrin 2 dynamics in a transiently transfected, apoptotic A431 cell. Images were obtained at 120 second intervals. Time-lapse sequence plays at 6 frames/second.

• Movie 4 -

  Movie 4. Time-lapse movie of apoptotic spike formation in UV-treated A431 cells. Images were obtained at 120 second intervals. Movie plays at 10 frames/second.

• Movie 5 -

  Movie 5. Effect of nocodazole on apoptotic spike formation in UV-treated A431 cells. Images were obtained at 120 second intervals. Movie plays at 10 frames/second.

• Movie 6 -

  Movie 6. Time-lapse movie of microtubule and chromatin dynamics throughout the duration of apoptosis. An anisomycin-treated A431 cell transiently-expressing YFP-tubulin (green) and HMGB1-CFP (red). AnnexinV labelling is depicted in blue. Images were obtained at 300 second intervals. Movie plays at 6 frames/second.

• Movie 7 -

  Movie 7. Time-lapse movie of EB1-GFP dynamics within an apoptotic spike of an A431 cell. The cell body is to the left. Confocal images were taken at 10 second intervals. Movie plays at 6 frames/second.