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. 2014 Apr 3;12(4):2054–2065. doi: 10.3390/md12042054

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© 2014 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/3.0/).

PMC Copyright notice

Effect of placotylene A (1) on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation. (A) Bone marrow-derived macrophages (BMMs) were cultured for 4 days with RANKL (10 ng/mL) and macrophage-colony stimulating factor (M-CSF) (30 ng/mL) in the presence of different concentrations of 1. The cells were fixed in 3.7% formalin, permeabilized in 0.1% Triton X-100, and stained for tartrate-resistant acid phosphatase (TRAP). (B) TRAP-positive cells were counted as osteoclasts (nuclei ≥3). ** P <0.01, *** P <0.001. (C) Cytotoxicity of 1 on BMMs. The effect of 1 on the viability on BMMs was evaluated using the CCK-8 assay.