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. 2014 Jun 5;289(31):21340–21350. doi: 10.1074/jbc.M114.550699

FIGURE 3.

FIGURE 3.

Ser-740 site of HDAC4 is important for dissociation from Runx2. A, mutation sites of GFP-rHDAC4; Serine residues mutated to alanine. B, UMR 106–01 cells were transfected with GFP-rHDAC4 or mutant GFP-rHDAC4 constructs. Cells were stimulated with PTH (1–34, 10−8 m) for 30 min. Total cellular lysates isolated from UMR 106-01 cells were immunoprecipitated with anti-Runx2 or control rabbit IgG antibodies. Samples were immunoblotted with anti-GFP antibody. The quantitative intensity was obtained using ImageJ. *, p < 0.05 versus respective control. C, UMR 106–01 cells were transfected with GFP rHDAC4 or mutant GFP rHDAC4 constructs (S355A, S576A, S740A, and T815A). Total cell lysates were immunoblotted with anti-GFP, anti-Runx2, and anti-Cdk antibodies. Anti-Cdk2 was used as a loading control.