Open Access Highly Accessed Research article

Human ASPM participates in spindle organisation, spindle orientation and cytokinesis

Julie Higgins1, Carol Midgley2, Anna-Maria Bergh3, Sandra M Bell1, Jonathan M Askham4, Emma Roberts1, Ruth K Binns1, Saghira M Sharif5, Christopher Bennett5, David M Glover3, C Geoffrey Woods6, Ewan E Morrison1 and Jacquelyn Bond1*

Author Affiliations

1 Section of Ophthalmology and Neuroscience, Wellcome Trust Brenner Building, Leeds Institute of Molecular Medicine, University of Leeds, St. James's University Hospital, Leeds LS9 7TF, UK

2 Department of Life Sciences, The Open University, Walton Hall, Milton Keynes, MK7 6AA, UK

3 Cancer Research UK Cell Cycle Genetics Research Group, University of Cambridge, Department of Genetics, Downing Street, Cambridge CB2 3EH, UK

4 CRUK Clinical Centre at Leeds, Division of Cancer Medicine Research, Leeds Institute of Molecular Medicine, St James's University Hospital, Leeds LS9 7TF, UK

5 Yorkshire Regional Genetics Service, Yorkshire Regional Genetics Service, Ashley Wing, St James's University Hospital, Leeds LS9 7TF, UK

6 Cambridge Institute for Medical Research, Wellcome Trust/MRC Building, Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2XY, UK

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BMC Cell Biology 2010, 11:85  doi:10.1186/1471-2121-11-85

Published: 2 November 2010

Additional files

Additional file 1:

A comparison of ASPM interphase localisation in HDF cells using ASPM N- and C-terminal ASPM antibodies. Cells were fixed and stained with the N-terminal ASPM antibody 216-1 or 217-2 or the C-terminal antibody 279-3 (green), anti-α-tubulin (red) and DAPI (blue) to identify nuclei. Scale bar = 10 μm.

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Additional file 2:

A comparison of ASPM metaphase localisation in HDF cells using ASPM N- and C-terminal ASPM antibodies. Cells were fixed and stained with the N-terminal ASPM antibody 216-1 or 217-2 or the C-terminal antibody 279-3 (green), anti-α-tubulin (red) and DAPI (blue) to identify nuclei. Scale bar = 10 μm.

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Additional file 3:

ASPM is localised at the spindle poles in metaphase cells for a range of cell types. HeLa, COS-7, U2OS, SH-SY5Y and HDF cells were fixed and stained with the N-terminal ASPM antibody 216-1 (green), anti-α-tubulin (red) and DAPI (blue) to identify nuclei. Scale bar = 10 μm.

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Additional file 4:

ASPM is positioned in a narrow ring at the centre of the midbody during telophase in a range of cell types. Cells were fixed and stained with the N-terminal ASPM antibody 279-3 (green), anti-α-tubulin (red) and DAPI (blue) to identify nuclei. Scale bar = 10 μm.

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Additional file 5:

Symmetrical division in GL3 luciferase control siRNA treated cells. A U2OS cell treated with GL3 siRNA undergoing symmetrical division with the mitotic spindle parallel to the surface of the imaging dish. This division results in two daughter cells and complete cytokinesis.

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Additional file 6:

Asymmetrical division and cytokinesis failure in a siRNA mediated ASPM depleted U2OS cell. A U2OS cell treated with ASP1 siRNA undergoing an asymmetric division with the mitotic spindle perpendicular to the surface of the imaging dish. This results in one daughter being extruded vertically from the dividing cell, towards the observer, while the other remains attached to the substrate. In this example cytokinesis fails, resulting in the formation of a binucleate cell.

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Additional file 7:

ASPM depleted binucleate U2OS cell undergoing apoptosis. This movie shows a binucleate U2OS cell following treatment with ASP1 siRNA, undergoing apoptosis.

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