Supplementary Materials

Microtubule detyrosination guides chromosomes during mitosis

Marin Barisic, Ricardo Silva e Sousa, Suvranta K. Tripathy, Maria M. Magiera, Anatoly V. Zaytsev, Ana L. Pereira, Carsten Janke, Ekaterina L. Grishchuk, Helder Maiato

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

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  • Materials and Methods
  • Figs. S1 to S12
  • Table S1
  • Full Reference List

Images, Video, and Other Other Media

Movie S1
Congression of pole-proximal chromosomes is impaired after CENP-E inhibition, TTL overexpression or TCP inhibition. Spinning-disk confocal time-lapse imaging of U2OS cells stably expressing H2B-GFP and mCherry-α-tubulin under the indicated experimental conditions. Cells were recorded every 2 min. Time=h:min.
Movie S2
Inhibition of tubulin carboxipeptidase with parthenolide dissociates CENP-E from microtubules in G2 cells. Spinning-disk confocal time-lapse imaging of HeLa cells stably expressing CENP-E-GFP and transfected with mCherry-α-tubulin under the indicated experimental conditions. Cells were recorded every 2 min. Time=h:min.
Movie S3
CENP-E motility on tyrosinated and detyrosinated microtubules in vitro. Single molecules of CENP-E-GFP (green) walk on taxol-stabilized microtubules that were polymerized from tyrosinated ("Tyr MT", red) or detyrosinated ("Det MT", purple) human tubulin. Video is played at 30 fps, which is 3 times faster than recorded.
Movie S4
Force generation by CENP-E motor. First image is an overlay of the DIC image of 0.54 *μm bead (grayscale), microtubule (red) and CENP-E-GFP (green); "+" symbol indicates microtubule polarity. All subsequent frames show motions of the bead (DIC) in a stationary laser trap. Images were acquired continuously every 10 ms, but the video shows only every 5th frame. Video is played at 30 fps, so the motions appear 1.4 times slower than recorded.
Movie S5
Chromosomes cannot complete congression in TTL depleted cells and are randomly transported away from the spindle poles by CENP-E. Spinning-disk confocal time-lapse imaging of U2OS cells stably expressing GFP-CENP-A and mCherry-α-tubulin under the indicated experimental conditions. Cells were recorded every 2 min. Time=h:min.