Fig. 4

Analysis of the regulatory role of TNF-α in LPS- induced osteoclastogenesis. a Measurement of TNF-α production in response to RANKL and LPS stimulation of RAW cells. ELISA determined the TNF-α concentrations in the culture medium. One-way ANOVA was applied, and values are expressed as mean ± standard deviation (SD). ***P < 0.001 vs. (−) control and RANKL- treated cells. b The diagrammatic sketch demonstrates the treatment strategy of RAW cells with LPS (5 μg/mL) and anti-TNF-α (2 μg/mL). c In response to the treatment strategy, representative images of TRAP stained osteoclasts are shown in panel B. TRAP stained osteoclasts in panels A and C were obtained with a 4X objective (magnification: 40X); panels in B and D were obtained with a 10× objective (magnification: 100X). d The number of TRAP-positive multinucleated osteoclasts were counted in both groups (n = 3). Statistical analysis was performed to compare the number of osteoclasts in the LPS+ anti- TNF-α group with the control group (LPS). The t-test was applied, and values are expressed as mean ± standard deviation (SD). **P < 0.01 vs. LPS group. LPS, lipopolysaccharide; RANKL, receptor activator of nuclear factor kappa-B ligand; TNF-α, tumor necrosis factor α