Detection of apoptosis by annexin V labeling in osteoclasts.

<p>(A) Osteoclasts were treated with CT (10 nM), SDCP (10 µM), or both for 18 h. Osteoclasts treated with TGF-βI (10 ng/mL) were included as positive controls. Green cellular membrane labeling indicates apoptotic cells. Only cells without propidium iodide (red) labeling were considered to be apoptotic. DIC images and nucleus stain were performed as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040272#pone-0040272-g001" target="_blank">Fig. 1</a>. Bar = 10 µm. (B) Quantitative results of the experiment shown in panel A. The measurement of apoptosis was calculated as the percentage of positive annexin V labeling cells in a total of at least 500 osteoclasts. <i>P</i><0.05 compared to *control group, †TGF-β1 group, ‡CT group; and §SDCP group after Bonferroni adjustment, mean ± SD, n = 6 in each group.