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. Author manuscript; available in PMC: 2014 Jun 1.
Published in final edited form as: J Bone Miner Res. 2013 Jun;28(6):1434–1445. doi: 10.1002/jbmr.1876

Figure 1.

Figure 1

Data generated from Western blot analysis showing OBs cultured alone, or OBs stimulated with MKs for 1, 2, and 4 hrs. MKs were removed (by washing) and OB lysates collected. A) Pyk2 expression in OBs was significantly increased with just 1 hr of co-culture with MKs and the increase persisted in OBs co-cultured for 2 and 4 hrs. B–D) OBs were pretreated with cycloheximide (Chx; 10 μM, optimal pretested) or actinomycin D (ActD; 5 μg/ml, optimal pretested) for 1 or 3 hrs, respectively. MKs were added to cultures as indicated and cells were incubated for 4 additional hrs (cultures containing inhibitors remained with inhibitors). B&C) Western blot analysis and D) Real-time PCR analysis. n=3/experimental group, representative data are shown from 3 independent experiments conducted (A–D). Pyk2 mRNA expression in OB and OB+MK cultures (real-time PCR data) was completed in more than 10 separate experiments with identical data generated from all of these experiments. *Indicates statistically significant differences (p<0.05) compared to 0 hr (A) or OB (B&C). ΨIndicates statistically significant difference (p<0.05) compared to OB+Chx (B) or OB+ActD (D). +Indicates statistically significant difference (p<0.05) compared to OB+MK (B–D).