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. 2017 Oct 25;159(1):272–284. doi: 10.1210/en.2017-00850

Figure 3.

Figure 3.

Temporal ablation of β-catenin expression reduces bone volume. (A) Schematic representation of experimental design. Eight-week-old mice were treated with TM (150 ng/g body weight) or sunflower seed oil (SSO) on 5 consecutive days and then euthanized at 9 weeks of age or at 17 weeks of age. (B and C) Immunofluorescent analysis of gene recombination in Oc-CreERT2; ROSAmT/mG treated with (B) SSO or (C) TM at 9 weeks of age (Original magnification ×4). (D–F) Immunofluorescent analysis of gene recombination in the trabecular bone compartment of ROSAmT/mG mice treated with (D) TM, Oc-CreERT2; ROSAmT/mG mice treated with (E) SSO, and Oc-CreERT2; (F) ROSAmT/mG mice treated with TM. (G–I) Immunofluorescent analysis of gene recombination in the (G) liver, (H) gonadal fat pad, and (I) skeletal muscle of Oc-CreERT2; ROSAmT/mG mice treated with TM. (J) Allele-specific PCR analysis of Ctnnb1 gene recombination in tissue isolated from Oc-CreERT2; Ctnnb1flox/flox mice (ΔCtnnb1) at 17 weeks of age (8 weeks after TM injection). (K) qPCR analysis of Ctnnb1 mRNA levels in the femur of control and ΔCtnnb1 mice. (L) Femur length and (M) growth plate length in control and ΔCtnnb1 mice at 17 weeks of age. (N) Representative computer renderings of trabecular bone structure in the distal femur (top) and L5 vertebrae (bottom) in control and ΔCtnnb1 mice. (O) Quantification of trabecular bone volume per tissue volume (BV/TV) in the distal femur and L5 vertebrae. (P) Quantification of trabecular bone number (Tb. N). (Q) Quantification of static and dynamic indices of bone turnover in 17-week-old control and ΔCtnnb1 mice. (R) qPCR analysis of phenotypic markers of osteoblastic cells in the femur of control and ΔCtnnb1 mice. (S and T) Serum levels of (S) P1NP and (T) CTX in control and ΔCtnnb1 mice. Data are expressed as mean ± standard error of the mean; n = 5 to 8 mice per group. *P < 0.05. BFR, bone formation rate; BPm, bone perimeter; BS, bone surface; MAR, mineral apposition rate; MS, mineralizing surface; NOb; osteoblast number; NOc, osteoclast number; OS, osteoid surface.