Fig. 2.

Reduced femur bone mass in TSK−/− mice. (A) Femurs were analysed separately in the trabecular region (cylinder: 0.4 mm proximal to the distal growth plate, 1.0 mm in height) and in the cortical region (cylinder: middle of the femur, 1.0 mm in height). (B) μCT images of femurs of 3-week-old WT and TSK−/− mice. (C) Three dimensional (3D) images of trabecular bones of 3-week-old WT and TSK−/− mice. (D) 3D images of cortical bones of 3-week-old WT and TSK−/− mice. (E, F) Parameters for the trabecular region, including bone volume/tissue volume ratio (BV/TV), trabecular number (Tb.N.), trabecular thickness (Tb.Th.), and trabecular separation (Tb.Sp.) in 3 and 20-week-old WT and TSK−/− mice. Data are presented as mean ± SD with 4 mice in each genotype group (*p < 0.05, **p < 0.01 vs. WT littermates, n = 4). (G) Cortical thickness (C.Th.) of 3- and 20-week-old WT and TSK−/− mice. C.Th. was measured at 8 points. Data are reported as mean ± SD (*p < 0.05 vs. WT littermates, n = 4). (H) Experimental outline of osteoblast differentiation. Bone marrow cells were harvested from femur of 20-week-old WT and TSK−/− mice and cultured with osteoinductive medium for 14 days and mineralized nodules were stained by alizarin red S. There was no significant difference in nodule formation activity between WT and TSK−/− osteoblasts. (I) Experimental outline of osteoclast differentiation. Bone marrow cells were harvested from femur of 20-week-old WT and TSK−/− mice and cultured with osteoclast-inductive medium for 7 days. And then, stained by tartrate-resistant acid phosphatase (TRAP) staining and the number of osteoclasts was counted. The number of TRAP-positive cells was significantly increased in TSK−/− mice.