Effects of PLE0 on osteoclast differentiation in vitro. (A,B) BMMs were cultured with vehicle (distilled water) or PLE0 (1.56–50 μg/mL) in the presence of M-CSF (60 ng/mL) and RANKL (100 ng/mL) for four days. (A) Osteoclast differentiation from BMMs in the presence or absence of PLE0 (magnification, ×100); (B) TRAP activity, number of cells, and viability of osteoclasts (Oc) after PLE0 treatment (0–200 μg/mL). After fixation and TRAP staining, TRAP-positive multinucleated giant cells (≥ three nuclei) were counted as osteoclasts. All data are represented as mean ± SEM and were analyzed with a one-way analysis of variance (ANOVA) and Dunnett’s post hoc test. ***, p < 0.001 versus no PLE0 treatment.