Figure 7

Combined effect of U0126 and LY294002 inhibited the osteoclast formation. (A) C7 cells were treated with 1 μM U0126 and 1 μM LY294002 for 24 hours. Cells were cultured in the presence of 25 ng/mL RANKL plus 50 ng/mL M-CSF. Cultures were fed every 3 days by replacing with 500 μL of fresh medium with or without minodronate, alendronate, U0126, LY294002, RANKL, and M-CSF. Cultures were fixed and stained for TRAP-positive multinucleated cells, and the number of cells per well was counted. *P < 0.01, compared to 25 ng/mL RANKL plus 50 ng/mL M-CSF administration. ^#P < 0.01, compared to 1 μM U0126 administration. (B) Cultures were fixed and stained for TRAP-positive cells, and the number of cells per well was counted. These results are representative of 4 independent experiments. *P < 0.01 compared to 25 ng/mL RANKL plus 50 ng/mL M-CSF administration.