Open Access Highly Accessed Research article

Multicellular tumor spheroid models to explore cell cycle checkpoints in 3D

Jennifer Laurent12, Céline Frongia12, Martine Cazales12, Odile Mondesert12, Bernard Ducommun123 and Valérie Lobjois124*

Author Affiliations

1 Université de Toulouse; ITAV-USR3505, F-31106, Toulouse, France

2 CNRS; ITAV-USR3505, F-31106, Toulouse, France

3 CHU de Toulouse, F-31059, Toulouse, France

4 ITAV - USR3505, Centre Pierre Potier, 1 place Pierre Potier, 50624, 31106, Toulouse Cedex 1, BP, France

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BMC Cancer 2013, 13:73  doi:10.1186/1471-2407-13-73

Published: 8 February 2013

Additional files

Additional file 1:

Characterization of Fucci-red and Fucci-green expressing spheroids. Immunostaining of Fucci-red and Fucci-green expressing cells with antibodies directed against cyclin D, cyclin E, cyclin A and cyclin B on small (A) or large (B) spheroids.

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Additional file 2:

Characterization of the effects of lovastatin and etoposide on Capan-2 cells lines expressing Fucci reporters. Capan-2 cell lines cultured as monolayers and treated or not with lovastatin (60 μM, 48 h) or etoposide (40 μM, 1 h). (A) Visualization of Fucci-red and Fucci-green expressing cells. (B) Flow cytometry analysis of DNA content after DRAQ5 staining. (C) Quantification of the percentage of Fucci-red and Fucci-green expressing cells.

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Additional file 3:

Etoposide treatment-induced DNA damages in MCTS. Capan-2 MCTS were treated or not with 5 μM etoposide for 48 h. The genotoxic effect of etoposide was analyzed by immunodetection of the phosphorylation of the H2AX histone variant (red) on frozen sections. Nuclei are stained with DAPI (blue). Scale bar: 50 μm.

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