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. 2010 Dec 18;286(11):9612–9622. doi: 10.1074/jbc.M110.174920

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FIGURE 4. — Inhibition of Hh signaling interferes with osteoclast differentiation and activity. A and C, inhibiting Hh signaling in the osteoclasts by the Smoothened (SMOH) inhibitor, cyclopamine (Cyclo, 20 μm), significantly (*, p < 0.0001) compromises their ability to differentiate (A) and resorb (C) matrix when stimulated with conditioned medium from breast cancer cells. RAW264.7 cells were cultured under differentiating conditions in the presence of breast cancer cell-conditioned medium/and cyclopamine (20 μm). Images were acquired at 10× magnification using the Nikon Eclipse TS 100 microscope. B and D, images represent differentiation assessed by TRAP staining (B) and resorption activity (D). In both B and D, the differentiation conditions included DM (panel a); DM + cyclopamine (panel b); and conditioned medium from SUM159 (panel c), MDA-MB-231 (panel e), and SUM1315 cells (panel g). Images in panels d, f, and h represent resorption in the presence of conditioned media from SUM159 (panel d), MDA-MB-231 (panel f), and SUM1315 cells (panel h) supplemented with cyclopamine.