Real-time RT-PCR analyses of the expression of osteocyte and osteoblast marker genes, Rankl, Opg, and Wnt antagonist genes at 4 months of age.
(A) Expression of Dmp1, Sost, Phex, Fgf23, and Mepe in the osteoblast fractions (OB, n = 6) and osteocyte fractions (OC, n = 9) of wild-type mice. The values of osteoblast fractions were defined as 1, and relative levels are shown. *vs. osteoblast fractions. (B) Expression of Dmp1, Sost, Phex, Fgf23, and Mepe in control groups of wild-type mice (n = 4) and BCL2 transgenic mice (n = 5). The values of wild-type mice were defined as 1, and relative levels are shown. (C) Expression of osteoblast marker genes. Expressions of keratocan (Kera), Runx2, Osterix, Col1a1, and osteocalcin (OCN) were examined by real-time RT-PCR using osteoblast-enriched samples and osteocyte-enriched samples from 7 wild-type mice at 6 weeks of age. The values of osteocyte fractions were defined as 1, and relative levels are shown. *vs. osteocyte fractions. (D) Rankl and Opg expression in osteoblast-enriched samples after unloading. Tail suspension was performed for 3 days using male wild-type mice [control group, 8 mice; unloaded group, 9 mice] and BCL2 transgenic mice [control group, 12 mice; unloaded group, 6 mice] at 4 months of age. (E) Expression of Rankl and Opg in the osteoblast fractions (OB) and osteocyte fractions (OC) from 6 and 11 wild-type mice at 4 months of age, respectively. The values of osteoblast fractions were defined as 1, and relative levels are shown. *vs. osteoblast fractions. (F) Expression of Rankl and Opg in osteocyte-enriched samples. Tail suspension was performed for 3 days using male wild-type mice [control group, 9 mice; unloaded group, 8 mice] and BCL2 transgenic mice [control group, 12 mice; unloaded group, 11 mice] at 4 months of age. (G) Expression of Wnt antagonist genes in osteocyte-enriched samples after unloading. Tail suspension was performed for 3 days using male wild-type mice and BCL2 transgenic mice at 4 months of age (8 mice in each group). The values of the control group of wild-type mice were defined as 1, and relative levels are shown in D, F, and G. In A–G, data are presented as the mean ± S.D. *vs. control in D, F, and G. *, ♯ P<0.05; **P<0.01; ***P<0.001.
