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. 2015 Aug 6;16(8):18293–18311. doi: 10.3390/ijms160818293

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© 2015 by the authors; licensee MDPI, Basel, Switzerland.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).

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Co-treatment with genistein (Gen) significantly suppressed MTX-induced ex vivo osteoclast formation. (A–C) Representative images (20×) of osteoclasts differentiated in the presence of macrophage colony stimulating factor (M-CSF) and receptor activator of NF-κB ligand (RANKL) from bone marrow mononuclear cells isolated from control, MTX-, and Gen + MTX-treated rats, respectively. Cells were subsequently stained with TRAP to identify osteoclasts; (D) Osteoclasts were identified as large, TRAP⁺, and multinucleated cells (≥3 nuclei) (black arrows in panels A–C) and quantified as osteoclasts/mm² of culture dish. n = 6, *** p < 0.001 vs. control.