Fig. 4

Osteoclastogenic activity is disrupted in Gli1-Cre^ERT2;Bmpr1α^fl/fl mice. a TRAP staining of the sagittal suture mesenchyme of Bmpr1α^fl/fl (control) and Gli1-Cre^ERT2;Bmpr1a^fl/fl (mutant) mice 1 month post induction (1mpt). The inset shows the boxed region magnified. Red arrows indicate mature osteoclasts. b Quantitation of TRAP-positive osteoclasts in the suture mesenchyme per section from three independent samples. c Immunostaining of RANK (green, indicated by arrows) in the suture mesenchyme of Bmpr1α^fl/fl (control) and Gli1-Cre^ERT2;Bmpr1α^fl/fl (mutant) mice 1 month post induction (1mpt). The inset shows the boxed region magnified. d Quantitation of RANK+ cells in the suture mesenchyme per section from three independent samples. e Real-time PCR of Tcirg1 in the suture mesenchyme from three independent Bmpr1a^fl/fl (control) and Gli1-Cre^ERT2;Bmpr1a^fl/fl (mutant) mice 2 weeks post induction. f Real-time PCR of RANKL and OPG and the ratio of RANKL/OPG in the suture mesenchyme from four independent Bmpr1α^fl/fl (control) and Gli1-Cre^ERT2;Bmpr1α^fl/fl (mutant) mice 1 month post induction. g TRAP staining of osteoclasts induced from BMMs of Bmpr1α^fl/fl (control) and Gli1-Cre^ERT2;Bmpr1α^fl/fl (mutant) mice 2 weeks post induction. The cells were cultured with M-CSF for 3 days and then with RANKL for another 5 days. The inset shows the boxed region magnified. Blue arrows indicate mature osteoclasts. h Quantitation of multinucleated TRAP-positive osteoclasts per well from four independent experiments. i Resorption activity assay (von Kossa staining) of osteoclasts induced from BMMs of Bmpr1α^fl/fl (control) and Gli1-Cre^ERT2;Bmpr1α^fl/fl (mutant) mice 2 weeks post induction, after culture with M-CSF for 3 days and then with RANKL for another 5 days. j Quantitation of resorption ratio per well from four independent experiments. T tests were performed. *P < 0.05; **P < 0.01. Broken lines indicate the outline of the suture. Scale bars, 100 µm