Fig. 5
The binding affinity on the promoters of ADAMTS genes was much higher for pRunx2S28 than for Runx2
(A) Greater amounts of p300 and NCOA3 protein were pulled down by pRunx2 than by Runx2. Equal weights (0.05 g) of three independent IVDs from LPS-challenged mice were mixed to make a homogenate, followed by immunoprecipitation with anti-Runx2-, anti-pRunx2S28, and IgG-coated protein A agarose. The purified complexes were used to determine the protein levels of Runx2, pRunx2S28, p300, and NCOA3. (B-F) Occupancies of Runx2 and pRunx2S28 on the promoters of ADAMTS1/2/3/4/5. The NP-1 cells were incubated with or without 20 ng/mL LPS for 6 h, followed by ChIP assays with anti-Runx2-, anti-pRunx2S28, and IgG-coated protein A agarose. The input and output DNA samples were used for RT-qPCR analyses to examine the occupancies of Runx2 and pRunx2S28 on the promoters of ADAMTS1(B), ADAMTS4(C), ADAMTS5(D), ADAMTS2(E), and ADAMTS3(F). *P < 0.05; ***P < 0.001; ns: no significant difference