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. 2013 Jul 25;8(7):e69468. doi: 10.1371/journal.pone.0069468

Figure 2. Effects of KMUP-1 on RANKL-induced osteoclastogenesis.

Figure 2

RAW264.7 cells were cultured with RANKL (10 ng/ml) for 5 days to induce osteoclast differentiation, with treatment of KMUP-1 (0–10 µM) to assess its anti-osteoclastogenic effects. In addition, cells treated with KMUP-1 (10 µM) had pretreatment of PKA inhibitor H89 (10 µM) and PKG inhibitor KT5823 (3 µM) to determine underlying mechanisms. (A) After 5 days of culture, TRAP positive cells and multinucleated osteoclasts were counted under the microscope. (B) The number of TRAP-positive multinucleated cells was counted. (C) TRAP activity was measured at 405 nm. Each value represents the mean ± S.E.M. of three independent experiments, with triplicate determinations in each experiment. ## P<0.01 compared with control; *P<0.05, **P<0.01 compared with RANKL alone; + P<0.05 compared with KMUP-1 10 µM plus RANKL.