Figure 2.

mTOR deficiency in osteoblasts resulted in reduced bone mass with impaired osteoblast differentiation. (a) Micro-CT images of femur from 4-week-old WT, mTOR^Osx/+ and mTOR^Osx littermates. (b–e) Quantitative parameters of Micro-CT. BV/TV: bone volume/total volume, Tb.N.: trabecular number, Tb.Th.: trabecular thickness, Ct.Th.: cortical thickness. Data represent means±s.d. *P<0.05, n=5. (f) Calvarial osteoblasts were cultured with osteogenic medium from 5-day-old WT and mTOR^Osx littermates. ALP staining and alizarin red staining were performed to analyze osteoblast differentiation on the 7th and 14th day respectively. (g) Immunohistochemistry for Runx2 of distal femur from 7-day-old WT and mTOR^Osx mice. (h) Number of Runx2-positive osteoblasts counted in the distal femur from 7-day-old WT and mTOR^Osx mice. Data represent means±s.d. *P<0.05, n=3. (i) In situ hybridizationfor Col1α1 and Ocn in femur from 7-day-old WT and mTOR^Osx littermaters. (j) Number of Col1α1-positive osteoblasts counted in the trabecular bone of distal femur from 7-day-old WT and mTOR^Osx littermaters. Data represent means±s.d. *P<0.05, n=3. (k) Number of Ocn-positive osteoblasts calculated in the cortical bone of distal femur from 7-day-old WT and mTOR^Osx littermaters. Data represent means±s.d. *P<0.05, n=3