Figure 6

LLDT-8 decreased the number of TRAP-positive cells. RAW264.7 cells were cultured in the presence of RANKL (50 ng/ml) and M-CSF (50 ng/ml) with various concentrations of LLDT-8 (0, 12.5, 25 and 50 nM, respectively) for 6 days, the cells of the control group were cultured without RANKL and M-CSF. All the cells were fixed and stained for TRAP according to the manufacturer’s instructions on day 1, 2, 3 and 6. Osteoclast formation was determined to be TRAP-positive staining multinuclear cells using light microscopy. The total number of TRAP-positive cells in each well was counted on day 1 (A), 2 (B), 3 (C) and 6 (D), respectively. #p < 0.05, ##p < 0.01, and ###p < 0.001, compared with the control group; *p < 0.05,**p < 0.01, compared with LLDT-8-non-treated (0 nM) group.