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. 2008 Jan 24;27(3):535–545. doi: 10.1038/sj.emboj.7601984

Figure 1.

Figure 1

Estrogen induces apoptosis in pre-osteoclasts and prevents the formation of mature osteoclasts. (A) Bone marrow was differentiated with M-CSF and RANKL for 8 days in the presence of ethanol (EtOH), 0.1 nM estradiol (E2), 1 nM E2 or 10 nM E2. Cells were then fixed and stained for TRAP. (B) The expression of cathepsin K mRNA or (C) calcitonin receptor mRNA was measured by quantitative real-time PCR from RNA isolated from bone marrow, pre-osteoclast and osteoclast cultures treated with or without 10 nM E2 for the entire course of differentiation. Fully differentiated osteoclasts treated with E2 were never formed and thus there is no value in this figure. The fold induction was normalized to β-actin mRNA. Error bars represent ±1 s.d. (D) Bone marrow was differentiated to the pre-osteoclast or osteoclast state with M-CSF and RANKL and then treated for 7 or 24 h with vehicle (EtOH), 1 nM E2 or 10 nM E2. Cells were fixed and apoptosis was detected by TUNEL. Quantification was performed using a laser scanning cytometer. Data are representative of three independent experiments.