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Open Access Research article

BMP2 and mechanical loading cooperatively regulate immediate early signalling events in the BMP pathway

Jessica Kopf12, Ansgar Petersen234, Georg N Duda234 and Petra Knaus1*

Author Affiliations

1 Institute for Chemistry/Biochemistry, FU Berlin, Berlin, Germany

2 Julius Wolff Institute, Charité-Universitätsmedizin Berlin, Germany

3 Berlin-Brandenburg Center for Regenerative Therapies, Berlin, Germany

4 CMSC, Charité-Universitätsmedizin, Berlin, Germany

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BMC Biology 2012, 10:37  doi:10.1186/1741-7007-10-37

Published: 30 April 2012

Additional files

Additional file 1:

BMP2 and mechanical loading synergistically regulate BMP-induced Smad phosphorylation events. (a and b) hFOBs were seeded on collagen scaffolds and subjected to BMP2 stimulation, mechanical loading or a combination of both. Protein lysates were analysed by western blot using specific antibodies.

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Additional file 2:

Smad2 is not phosphorylated by BMP2 stimulation, mechanical loading or a combination of both. hFOBs were seeded on collagen scaffolds and subjected to BMP2 stimulation, mechanical loading or a combination of both for indicated time points. As positive control, hFOBs were stimulated for 30 minutes with 100 pM TGF-β1. Protein lysates were analysed by western blot using specific antibodies.

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Open Data

Additional file 3:

BMP2 and mechanical load synergistically regulate Id1 gene expression in primary human mesenchymal stem cells. Data of one representative experiment is depicted. Human primary mesenchymal stem cells (hMSCs) were embedded in fibrin gels and loaded for 3 days in the absence or presence of 10 nM BMP2. Embedding and loading was performed as described previously [68]. Total RNA was extracted and Id1 gene expression was analysed by qRT-PCR.

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Additional file 4:

Mechanical loading protocol. Running protocol for short-term (up to 120 minutes) and long-term (up to 24 hours) mechanical loading.

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Additional file 5:

Primer sequences. Sequence of primers used for qRT-PCR.

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Additional file 6:

Validation of a reference gene for qRT-PCR. Validation of HPRT as house-keeping reference gene using geNorm software [69].

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Open Data