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. 2010 Sep 17;285(47):36410–36419. doi: 10.1074/jbc.M110.142307

FIGURE 6.

FIGURE 6.

Erk activation increases the association between Runx2, Smad1 and p300. A, C2C12 cells were transiently transfected with Myc-Runx2, FLAG-Smad1, and HA-p300 expression vectors and incubated for 24 h in the presence or absence of BMP-2 (200 ng/ml) and/or U0126 (40 μm). IP and/or IB analysis was then performed. Densitometry analysis results were provided under the respective band. B, 293T cells were transiently transfected with Myc-Runx2, FLAG-Smad1, HA-p300, and MEK1 expression vectors as indicated and incubated for 24 h. IP and/or IB analysis was then performed. C and D, S310A/S319A mutation reduces Erk activation-induced Runx2 acetylation and stabilization. C, 293T cells were transiently transfected with wild type Myc-Runx2 (WT) or Myc-Runx2-S301A/S319A (SA-301/319) concomitantly with FLAG-Smad1, HA-p300, and MEK1 expression vectors as indicated and incubated for 24 h. IP and/or IB analysis was then performed. AcK, anti-acetylated lysine antibody, P-S, anti-phosphoserine antibody. D, C2C12 cells were transiently transfected with WT or SA mutant Myc-Runx2 expression plasmids and were treated with BMP-2 (80 ng/ml) for 18 h as indicated. IP and/or IB analysis was then performed.