Fisetin Inhibits Osteoclast Differentiation via Downregulation of p38 and c-Fos-NFATc1 Signaling Pathways
Figure 3
NFATc1 over-expression restores fisetin-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 (5 ng) for 4 days in the presence or absence of fisetin (5 μM). After 4 days, cells were fixed and GFP expression was visualized under a fluorescence microscope. (b) BMMs were infected with GFP or CA-NFATc1-GFP and then 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.
