Additional file 1.

M-CSF and RANKL-induced monocyte-to-osteoclast differentiation. (A) Visualization of the formation of the actin ring and the generation of polykaryons in monocyte (MO) to osteoclast (OC) differentiation with phalloidin and DAPI staining. (B) TRAP (Tartrate resistant acid phosphatase-OC marker) staining in MO and OC preparations, showing this activity only in OCs. Determination of the typical percentage of osteoclastic nuclei present in the preparations used for the experiments; over 84% efficiency was achieved at 21 days. (C) Upregulation of OC specific markers (CA2, CTSK, MMP9, ACP5) was checked by qPCR; downregulation of a monocyte specific gene (CX3CR1) was also monitored.

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de la Rica et al. Genome Biology 2013 14:R99   doi:10.1186/gb-2013-14-9-r99