Figure 6. Reduced number of TRACP-positive multinuclear cells (osteoclasts) in human peripheral blood monocyte cultures subjected to conditioned medium from RAD001- or mevinolin-treated R527H LMNA U2-OS cells.

(A) Human mononuclear cells were induced to differentiate into osteoclasts for 8 days by adding medium from untreated WT LMNA, R527H LMNA U2-OS, or R527H LMNA U2-OS treated with RAD001 (72 hours) or mevinolin (24 hours). TRACP expression is revealed by red precipitates in representative pictures, nuclei are counterstained with Hoechst 33258 (Bar, 20 μm). Arrowheads indicate osteoclasts and their nuclei. (B) Quantitative analysis of osteoclast formation. Osteoclast number per sample is reported on the Y axis. Analysis was performed on six countings of the TRACP positive cells that showed three or more than three nuclei. Data are expressed as mean+/− standard deviation. Statistically significant differences (p < 0.05) are indicated by asterisks.