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Files in this Data Supplement:
Fig. S1. Examples of Ptk1 ana-telophases after M-FISH showing the external localization of chromosome 1 with the chromosome arms protruding towards the periphery of the group of migrated chromosomes.
Fig. S2. Mitotic spindle reorganization after NOC or MON treatment. (A) Mitotic spindle reassembly as visualized by centrosome (γ-tubulin, green) and microtubule (α-tubulin, red) staining at different times after NOC washout in PtK1 cells. Extra-centrosomal foci of α-tubulin, promoting merotelic orientation of kinetochores, were present at early times during spindle reassembly. Time is in minutes. Scale bar: 10 µm. (B) Chromosome dynamics at different times from NOC release as observed by DAPI staining: prometaphase (one or more unaligned chromosomes), metaphase (complete chromosome alignment), ana-telophase (after chromatid separation); time in minutes. (C) Centrosome dynamics at different times from NOC release as observed by γ-tubulin staining: monopolar (two adjacent γ-tubulin signals), intermediate (intermediate distance of the two γ-tubulin signals), bipolar (the two γ-tubulin signals lay at the opposite sites of the cell). (D) Mitotic spindle reassembly in cells fixed at different time after MON washout. Markers are described in A. Time is in minutes. Scale bar: 10 µm. (E) Chromosome and (F) centrosome dynamics after MON treatment. Categories are described in B and C. 100-300 cells in three independent experiments were counted for each time point.
Fig. S3. Distribution of the different mitotic phases in PtK1 cells as assayed by DAPI staining. Chromosome dynamics was analyzed in untreated cells (DMSO), at the end of the NOC treatment (NOC) or in cells released for 15 minutes in drug-free medium (NOC+15) or released and incubated in MG132 for 1 hour (NOC+15+1h MG132) or 2 hours (NOC+15+1h MG132). After 1 hour MG132 treatment the percentage of prometaphase cells was low enough to guarantee that the hesperadin treatment was performed on cells in which syntelic orientations were already corrected. 300-700 cells from three to six independent experiments were counted for each time point.
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