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. 2007 Feb 26;176(5):709–718. doi: 10.1083/jcb.200610046

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Copyright © 2007, The Rockefeller University Press

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Figure 3. — Changes in RUNX2 phosphorylation and transcriptional activity in osteoblasts from TgMek-dn and -sp mice. (a and b) Regulation of RUNX2 phosphorylation. Calvarial cells were grown under differentiating conditions for 7 d before metabolic labeling with [³²P]orthophosphate or [³⁵S]methionine/cysteine (to normalize for total RUNX2) and immunoprecipitation with an anti-Runx2 antibody. Each IP reaction contained 500 μg total protein. (b) Normalized ³²P incorporation into RUNX2. (c and d) Runx2-dependent transcriptional activity. Cells were transfected with p1.3mOG2-luc (c) or p6OSE2mOG2-luc (d) plasmids and grown under differentiating conditions for the times indicated before measurement of luciferase activity. Values are the means ± the SD of triplicate independent samples.