Figure 1
From: Msx2 mediates the inhibitory action of TNF-α on osteoblast differentiation
TNF-α suppresses BMP2-induced ALP expression in Runx2-/- cells. (A, B) C2C12 cells were incubated in DMEM supplemented with 5% FBS for 24 h in the presence of the indicated reagents. Runx2+/+ and Runx2-/- cells were incubated in α-MEM supplemented with 10% FBS, 10 mM β-glycerophosphate and 50 µg/ml ascorbic acid for 48 h in the presence of the indicated reagents. Then, ALP staining (A) or RT-PCR (B) was performed. (C) To block new protein synthesis, C2C12 cells were treated with BMP2 and/or TNF-α in the presence of cycloheximide (CHX, 10 µg/ml) for 24 h. The concentrations of TNF-α and BMP2 were 10 ng/ml and 100 ng/ml, respectively, or as otherwise indicated. *, In Runx2-/- cells, the RT-PCR products of the ALP gene were obtained by carrying out the amplification step for five more cycles than in C2C12 and Runx2+/+ cells.
