Figure 6. Olfm2 physically interacted with Runx2 to inhibit Myocd-SRF binding.

A and C, Olfm2 bound to Runx2. RASMCs were treated with vehicle (−) or PDGF-BB (+, 20 ng/ml) for 24 h followed by Co-IP with Olfm2 antibody (A) and by the reverse Co-IP with Runx2 antibody (C). The immunoprecipitated lysates were immunoblotted (IB) with the antibodies as indicated. B and D, Quantification of Olfm2-bound Runx2 (B) and Runx2-bound Olfm2 (D) by normalizing to the corresponding TUBA1 level shown in A and C, respectively *, P < 0.01 vs. corresponding vehicle-treated group (−) (n=3). E, Knockdown of Runx2 restored Olfm2-inhibited SRF-Myocd binding. Ad-GFP- or Ad-shRunx2-transduced RASMCs were transfected with Olfm2 expression plasmid followed by Co-IP with IgG or SRF antibody. The immunoprecipitated lysates were immunoblotted (IB) with the antibodies as indicated. F–H, Quantification of SRF-bound proteins as indicated in E by normalizing to the input SRF level. *, P < 0.01 vs. Ad-GFP group without Olfm2; #, P < 0.01 vs. Ad-GFP group with Olfm2 transfection (n=3).