Figure 3

NFATc1 overexpression restores matairesinol-mediated inhibition of osteoclast differentiation. (A) BMMs were infected with pMX-IRES-GFP (GFP) or pMX-IRES-CA-NFATc1-GFP (CA-NFATc1-GFP) for 8 h with polybrene (10 μg/ml). Infected BMMs were cultured with M-CSF (30 ng/ml) and RANKL (10 ng/ml) for 4 days in the presence or absence of matairesinol (10 μM). After 4 days, cells were fixed and GFP expression visualized under a fluorescence microscope. (B) BMMs were infected with GFP or CA-NFATc1-GFP, and cells were cultured as described in (A). After 4 days, mature TRAP-positive multinucleated osteoclasts were visualized by TRAP staining. (C) TRAP-positive cells were counted as osteoclasts. (D) TRAP activity was measured at 405 nm.