A-C. BMCs derived from Fbw7 cKO or control mice were cultured in the presence of M-CSF (50 ng/mL) for 3 days for differentiation into BMMs, and further cultured with 100 ng/mL RANKL for 3 days for differentiation into osteoclasts. Cells were fixed and stained for TRAP. At 3 days prior to fixation, cells were treated with increasing concentration of DAPT (A), GSI-1 (B), or zoledronic acid (C). The TRAP-positive MNCs, which display cytoplasmic red staining and a minimum of three nuclei, were counted as osteoclasts. Data represents mean ± SD, n = 3, **p < 0.01, Student’s t test.
D. Micro-CT images of the distal femurs derived from Fbw7 cKO or control mice that were intraperitoneally injected with DMSO, DAPT (5 mg/kg), GSI-1 (5 mg/kg), or zoledronic acid (ZOL) (250 μg/kg) every four days for 8 weeks. Scale bar: 1 mm.
E. Microstructural parameters of the distal femurs analyzed in (D). Data represents mean ± SD, n = 5, *p < 0.05, Student’s t test.
See also Figure S7