1. Fig. 5. Recovery steps of mitotic acentriolar cells after cold MT depolymerisation.(A,B) Time-lapse sequences of mitotic spindle re-formation at 18°C.
Recovery steps of mitotic acentriolar cells after cold MT depolymerisation.(A,B) Time-lapse sequences of mitotic spindle re-formation at 18°C. Time 0 corresponds to the first image recorded on the microscope (see Materials and Methods). (A) WT cell (Jupiter line). MTs polymerisation occurs essentially from the centrosomes and spindle forms under an “outside-in” process. Centrosomes were first close to the chromosomes and then move away. Scale bar: 5 µm. (B) Acentriolar cell (line 131). Spindle forms from the chromosomes under an “inside-out” mechanism. Scale bar: 5 µm. (C) Immunofluorescence images of acentriolar cells fixed at various times of recovery at 23°C. (a) after 3 minutes of recovery at 23°C, multiples dots of nascent MTs are first observed in the immediate vicinity of the chromosomes. (b) after 6 minutes of recovery elongated bundles of MTs are observed from discrete sites onto the chromosomes. (c) these sites are identified by CID protein as the kinetochores. Inlay: magnified view of c. Blue: DNA, Green: α-tubulin, Red: CID. Scale bar: 10 µm.
Nicolas Lecland et al. Biology Open 2012;bio
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