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. 2017 Jul 21;158(11):3778–3791. doi: 10.1210/en.2017-00159

Figure 5.

Figure 5.

MEF2C associates with c-FOS on the Mmp13 promoter after PTH treatment in osteoblastic UMR 106-01 cells. (A) UMR 106-01 cells were treated with or without 10−8 M PTH-(1-34) for 1 hour, and lysates were immunoprecipitated (IP) with anti-MEF2C, c-FOS, c-JUN, or normal IgG (Santa Cruz), then immunoblotted with anti-MEF2C-FOS, c-JUN. (B) UMR 106-01 cells were treated with or without PTH for the indicated times and collected for ChIP assays with antibodies against c-FOS, or IgG as a negative control. DNA was amplified by real-time PCR using primers for the rat Mmp13 proximal promoter region. Data shown are mean ± SE. *P < 0.05, **P < 0.01, ***P < 0.001 vs control in each set, n = 3. (C) UMR 106-01 cells were treated with or without PTH for 4 hours. Cell lysates were collected for ChIP-reChIP assays with antibodies against MEF2C or IgG as a negative control for the first ChIP, and antibodies to c-FOS or IgG were used for the second immunoprecipitation. DNA was amplified by real-time PCR using primers for the rat Mmp13 proximal promoter region. Data shown are mean ± SE. ***P < 0.001 vs control, n = 3. (D) Cell lysates were collected for ChIP-reChIP assays with antibodies against MEF2C or IgG as a negative control for the first ChIP, and antibodies to c-JUN or IgG were used for the second immunoprecipitation. (E) Cell lysates were collected for ChIP-reChIP assays with antibodies against c-FOS or IgG as a negative control for the first ChIP, and antibodies to c-JUN or IgG were used for the second immunoprecipitation. Data shown are mean ± SE. *P < 0.05 vs control, n = 3.