Fig. 3

Portulaca oleracea ethanol extract (POEE) markedly enhances receptor activator of NF-κB ligand (RANKL)-mediated tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cell (MNC) formation and TRAP activity. a and b Isolated bone marrow-derived macrophages (BMMs) were simultaneously treated with RANKL (50 ng/mL) and POEE (0, 25, 50, and 100 μg/mL) and then cultured for 4 d. Following incubation, TRAP staining was performed. TRAP-positive MNCs present in each well, identified by the presence of more than 3 nuclei and cell size larger than 100 μm in diameter, were counted. c To measure TRAP activity, cells were treated with RANKL (50 ng/mL) and POEE (50 μg/mL) simultaneously. After 4 d of incubation, cells were lysed and total TRAP activity was then measured as described in “Materials and Methods”