Effect of RUNX1 on BMP9-induced classical Smad1/5/8 signaling and MAPKs (Mitogen-activated protein kinases) signaling. Cells were infected with Ad-RUNX1, Ad-siRUNX1, and Ad-BMP9, respectively. Total protein was extracted and was measured by Western blot. (A) Ad-BMP9, Ad-RUNX1 and Ad-siRUNX1 could infect MEFs cell; W (white light), F (fluorescence). (B,C) Effect of RUNX1 on BMP9-activated Smad1/5/8 signaling. (D,E) Effect of RUNX1 on BMP9-induced p38 and ERK1/2 (Extracellular signal-regulated protein kinase1/2) signaling. (F) A schematic presentation of proposed mechanism of BMP9/RUNX1 regulation of osteogenesis, P (phosphorylation), black arrow (Other studies confirm the phenomenon), red arrow and curved red arrow (our study confirm the phenomenon). * p < 0.05, as compared with BMP9 group. Scale bar represents 50 μm Magnification, ×100.