Barley Seedling Extracts Inhibit RANKL-Induced Differentiation, Fusion, and Maturation of Osteoclasts in the Early-to-Late Stages of Osteoclastogenesis

Figure 4

BSE also inhibits RANKL-mediated cell fusion during osteoclastogenesis. (a) Based on the exposure schedule, the BMMs were cultured with BSE (3 ug/ml) for various times periods (the indicated black arrow) in the presence of M-CSF (30 ng/ml) and RANKL (10 ng/ml). (b) After the BMMs differentiated into osteoclasts (as described in (a)), the cells were fixed, permeabilised, and stained with TRAP. TRAP+ MNCs formation was photographed under a light microscope. Each exposure period of BSE was indicated as “0–4” for the vehicle. “0-1” for 0-1 day, “1-2” for 1-2 days, “2-3” for 2-3 days, and “3-4” for 3-4 days. (c) TRAP activity was measured at 405 nm. ^### (versus the control); (versus the RANKL-treated control). (d) The number of TRAP+ MNCs (nuclei > 10) was counted. ; (versus the RANKL-treated group). (e) The effect of BSE on the mRNA expression of DC-STAMP was analysed using real-time PCR, as described in Figure 2(a). HPRT was used as the internal control. White column (vehicle-treated); black column (3 μg/ml BSE-treated), (versus the vehicle control). Data are representative of at least three experiments.

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