Fig. 1

Osteoclastogenic potential of BC cells and effects of Everolimus. PBMCs from healthy donors were differentiated to OCs using osteoclastogenic factors (left) and conditioned medium (CM) from MDA-MB-231 and MCF-7 BC cell lines, before (middle) and after (right) treatment with Everolimus. The pro-OC potential was measured as numbers of both large multinucleated OC-like cells stained by hematoxylin/eosin (a) and multinucleated TRAcP⁺ cells (b). CM from BC cells produced significant increases (*p < 0.05) of OC formation, although the greatest effect was with MCF-7 cells (middle). Both constitutive and BC-mediated osteoclastogenesis were abrogated when using CM from BC cells pretreated with a sub-lethal dose of Everolimus. The pattern of increased/reduced OC formation was similarly evident after different staining. a bottom: Polykaryons from PBMCs cultured with RANKL/M-CSF (left), with CM from untreated (middle) and Everolimus-treated (right) MCF-7 cells (scale bar: 30 μm; 30X magnification). b bottom: images of TRAcP⁺ OC-like cells (scale bar: 35 μm; 20x magnification)