Osteoclast differentiation in cultures of bone marrow cells from WT and <i>Gli1</i>^+/− mice.

<p>(<b>A</b>) Formation of TRAP-positive multinucleated osteoclasts by the co-culture of BMSCs and BMMΦ derived from WT or <i>Gli1</i>^+/− mice. (<b>B</b>) The numbers of osteoclasts expressed as means ± SDs of 4 wells per group in (A). *p<0.05 vs. the control group (WT BMSC × WT BMMΦ). (<b>C</b>) mRNA expression of <i>Rankl</i>, <i>Opg</i>, <i>Dmp1</i>, and <i>Sost</i> in 14-day osteogenic cultures of BMSCs. The mRNA expression was analyzed by real-time RT-PCR. <i>Rankl</i>, receptor activator of nuclear factor-κB ligand; <i>Opg</i>, osteoprotegerin; <i>Dmp1</i>, dentin matrix acidic phosphoprotein 1; <i>Sost</i>, sclerostin. *p<0.05 vs. WT. (<b>D</b>) Formation of TRAP-positive multinucleated osteoclasts in 5-day cultures of RAW cells in the presence or absence of sonic hedgehog (Shh) and cyclopamine (Cyc). (<b>E</b>) The numbers of osteoclasts in (D) expressed as means ± SDs of 4 wells per group. (<b>F</b>) mRNA expression of <i>Ptch1</i> in cultured RAW cells in the presence or absence of Shh and cyclopamine (Cyc). Data are means ± SDs of 4 wells per group. (<b>G</b>) mRNA expression of cathepsin K (<i>Ctsk</i>) and <i>NFATc1</i> in osteoclasts derived from WT or <i>Gli1</i>^+/− BMMΦ. Cells were cultured with recombinant M-CSF (10 ng/mL), RANKL (100 ng/mL), and Shh (25 nM). The mRNA expression was analyzed by real-time RT-PCR.