Research article
Gene expression profiling of early intervertebral disc degeneration reveals a down-regulation of canonical Wnt signaling and caveolin-1 expression: implications for development of regenerative strategies
1 Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 108, 3508 TD, Utrecht, The Netherlands
2 Department of Clinical Sciences, Division of Small Animals, Faculty of Veterinary Medicine and Animal Sciences, Swedish University of Agricultural Sciences, Ulls väg 14 C, SE-75651 Uppsala, Sweden
3 Department of Biochemistry and Cell Biology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL, Utrecht, the Netherlands
4 Department of Pathobiology, Pathology Division, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 108, 3508 TD, Utrecht, The Netherlands
5 Molecular Cancer Research, University Medical Center Utrecht, Utrecht, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands
6 Department of Orthopedics, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX, Utrecht, The Netherlands
Arthritis Research & Therapy 2013, 15:R23 doi:10.1186/ar4157
See related editorial by Erwin, http://arthritis-research.com/content/15/2/113
Published: 29 January 2013Additional files
Additional file 1:
Table S1 Primers used for qPCR analysis. Table describing the complete primer information for all target and reference genes investigated.
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Additional file 2:
Data analysis and statistics. Complete description of the data analysis and statistics used for the microarray analyses and the qPCR, immunohistochemistry, and immunofluorescence analyses.
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Additional file 3:
Table S2 N-fold changes and P-values for microarray and qPCR analysis. N-fold changes and corresponding P-values for micorarray and qPCR analysis of notochordal marker genes, caveolins, and genes involved in canonical Wnt signaling.
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Additional file 4:
Table S3 Top regulated genes for microarray analysis. Results obtained for the microarray comparisons between the notochordal cell (NC)-rich nucleus pulposus (NP), mixed NP, and chondrocyte-like cell (CLC)-rich NP in non-chondrodystrophic and chondrodystrophic dogs, and between the breed types for each histological stage.
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Additional file 5:
Table S4 Top five most significantly regulated pathways. Top five most significantly regulated pathways on the basis of all performed microarray comparisons using Metacore pathway analysis.
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Additional file 6:
Figure S1 Beta-catenin protein expression in the chondrocyte-like cell (CLC)-rich nucleus pulposus (NP) of non-chondrodystrophic and chondrodystrophic dogs. Immunohistochemistry (typical examples and quantification of expression) and western blot analysis for β-catenin protein expression in the chondrocyte-like cell (CLC)-rich nucleus pulposus (NP).
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Additional file 7:
Figure legends for Additional files 6and 9. Figure legends for Additional file 6, Figure S1 and Additional file 9, Figure S2.
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Additional file 8:
Table S5 Linear mixed model results for Caveolin-1 immunohistochemistry. Linear mixed model results for the Caveolin-1 immunohistochemistry analyses of healthy vs. early-degenerated nuclei pulposi in dogs with naturally occurring intervertebral disc degeneration.
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Additional file 9:
Figure S2 Caveolin-1 gene and protein expression in primary notochordal cells in monolayer culture. Caveolin-1 gene and protein expression in primary notochordal cells in monolayer culture.
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Additional file 10:
Table S6 P-values for mixed model analaysis of Caveolin-1 expression in culture. P-values for the mixed model analyses of Caveolin-1 gene and protein expression of notochordal cells in culture.
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