Figure 2. Low bone formation in FoxO1_ob-/- mice.

A) Bone mineral density (BMD) measured by DEXA in spine and femur of 2 month-old WT, FoxO1_ob+/- and FoxO1_ob-/- mice (n=10 mice/group). * p < 0.05 vs WT.

B) BV/TV, bone volume over trabecular volume; N.Ob/T.Ar, number of osteoblasts per trabecular area; BFR, bone formation rate, of 2 month-old and OcS/BS, osteoclast surface per bone surface in verterbrae of 1 month-old WT and FoxO1_ob-/- mice (n=10 mice/group). * p < 0.05 vs WT.

C) BV/TV in femoral head and midshaft cortical thickness in the femurs of 2 month-old WT and FoxO1_ob-/- mice (n=10 mice/group). * p < 0.05 vs WT.

D) BV/TV, BFR and N.Ob/T.Ar in verterbrae of 6 and 12 month-old WT and FoxO1_ob-/- mice (n=5-10 mice/group). * p < 0.05 vs WT.

E, F) BMD measured by DEXA in spine and femur of 2 month-old WT and FoxO3_ob-/- mice (n=10 mice/group).

G) Immunoblotting analysis of FoxO4 activity in the bone of WT and FoxO1_ob-/- mice.

H) RT-PCR analysis of FoxO4 expression in osteoblasts transfected with siRNA oligos for FoxO4 or control (Ctrl), scrambled oligos. * p < 0.05 vs control (Ctrl) siRNA and Ctrl siRNA in OM. OM denotes Osteogenic Medium.

I) Proliferation in cultures of osteoblasts transfected with siRNA oligos for FoxO4 or control (Ctrl), scrambled oligos. OM denotes Osteogenic Medium.

J) Real-time PCR (RT-PCR) analysis of CyclinD1, D2 and p27Kip1 expression in osteoblasts with silenced FoxO4.

K) RT-PCR analysis of osteoblast differentiation markers in osteoblasts with silenced FoxO4. In all panels values are means ± sem.