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. Author manuscript; available in PMC: 2016 Nov 21.
Published in final edited form as: J Bone Miner Res. 2014 Nov;29(11):2427–2438. doi: 10.1002/jbmr.2282

Figure 4. Disruption of IGFBP-2/RPTPβ interaction impairs IGF-I-stimulated AKT activation and osteoblast differentiation.

Figure 4

(A) Lysates from quiescent LacZ or IGFBP-2 overexpressing cells obtained on day 6 after differentiation medium (DM) exposure treated with or without IGF-I alone (10 min) or IGF-I following a 4 hr exposure to anti-fibronectin domain (FN3) antibody were immunoprecipitated using an anti-IGFBP-2 antibody and immunoblotted with an anti-RPTPβ antibody (B) Lysates from quiescent IGFBP-2 overexpressing cells that received the same treatments as in panel A were obtained on Day 6 after DM exposure and were immunoblotted with indicated antibody. (C) Lysates from LacZ overexpressing cells obtained on day 21 after DM exposure following incubation with the indicated concentration of anti-fibronectin antibody (FN3) were immunoblotted with anti-osteocalcin and anti-β-actin antibodies. (D) Lysates from IGFBP-2 overexpressing cells obtained on day 9 and 12 after DM exposure following incubation with PQ401 (10 uM) were immunoblotted with anti-osteocalcin and anti-β-actin antibodies. (E) Cells expressing IGFBP-2 that had been incubated with or without PQ401 were stained with Alizarin Red on day 18 after DM exposure.

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