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Figure 3 | BMC Biology

Figure 3

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

Figure 3

Bone morphogenetic protein 2 and mechanical loading synergistically regulate bone morphogenetic protein-induced Smad phosphorylation events. (a) 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. (b) Quantification of western blot analysis depicting phosphoprotein levels normalized to GAPDH. Bar charts represent means ± standard error of the mean (SEM) from three independent experiments; *P < 0.05; **P < 0.01. (c) hFOBs were seeded on collagen scaffolds and subjected for 90 minutes to BMP2 stimulation, mechanical loading or a combination of both. Gene expression was analysed by qRT-PCR. Bar charts summarize three independent experiments and depict means ± SEM. (d) hFOBs were seeded on collagen scaffolds and subjected for 30 min to BMP2 stimulation, mechanical loading or a combination of both. Nuclear and cytosolic protein lysates were fractionated and analysed by western blot. BMP: bone morphogenetic protein; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hFOB: human fetal osteoblast; qRT-PCR: quantitative reverse transcriptase polymerase chain reaction; SEM: standard error of the mean.

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