IAP antagonists enhance osteoclastogenesis in vitro.

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IAP antagonists enhance osteoclastogenesis in vitro. IAP antagonists enhance osteoclastogenesis in vitro. A, BMMs were cultured with indicated doses of BV6, 20 ng/mL GST-RANKL plus vehicle (Veh), or BV6 plus RANKL for 6 days, then visualized by TRAP staining. BV6 and vehicle were given in 2-hour pulses every other day, and RANKL was always present. TRAP-positive multinucleated cells were counted in each well and plotted. mOC, mouse osteoclasts. B, BMMs were cultured as in A, and RNA was collected on days 0, 2, and 4 for real-time reverse transcriptase PCR (RT-PCR) analysis of osteoclast differentiation markers. Data are plotted as fold change relative to day 0. C, mouse BMMs were treated with constant 52S (0.3 μmol/L) during osteoclast differentiation. mOC per well were counted and plotted. D and E, human peripheral mononuclear cells were cultured in osteoclastogenic media and pulse treated with BV6 (D) or 52S (E), as described earlier. Mature osteoclasts (hOC) were TRAP stained and counted. Scale bars, 1 mm. Data represent the mean ± SD; *, P < 0.05; **, P < 0.01; ***, P < 0.001. Chang Yang et al. Cancer Discovery 2013;3:212-223 ©2013 by American Association for Cancer Research