Figure S1. Increasing insulin signaling in osteoblasts...
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Figure S1. Increasing insulin signaling in osteoblasts weakens glucose intolerance in mice fed a HFD. (A) Histomorphometric analysis of vertebrae of WT and α1(I)Col-InsR mice fed a ND (n=8), (B) Serum Ctx levels in WT and α1(I)Col-InsR mice fed a ND, (C) Serum osteocalcin levels in WT and α1(I)Col-InsR mice fed a ND (n=8), (D) qPCR analysis of the expression of Glut1, Glut3 and Glut4 in bones of WT and α1(I)Col-InsR fed a ND or HFD (n=5). (E) Western blot analysis of the phosphorylation levels of the INSR and AKT in liver and WAT of WT and α1(I)Col-InsR mice fed a ND or a HFD. All Western blot experiments were repeated at least three times.*P<0.05. WT ND Col1a1-INSR ND A B 0 Expression relative to control (fold) 0.5 1.0 1.5 2.0 Glut1 Glut3 Glut4 WT HFD C Osteocalcin level(ng/ml) 50 200 0 100 150 Total-Ocn Glu13-Ocn 250 300 D INSR P-INSR (Y1150/Y1151) `<ACTIN P-AKT (S473) AKT Insulin: 0.5u/kg - + - + - + - + ND HFD ND HFD Liver - + - + - + - + ND HFD ND HFD WAT WT WT E * * BV/TV (%) 5 10 0 15 Oc.S/BS (%) 5 10 0 15 20 BFR/BS (+m 3 /+m 2 /y) 0 50 100 150 200 Serum Ctx(ng/ml) 5 10 0 15 20 25 Tg WT ND Col1a1-INSR ND Tg WT ND Col1a1-INSR ND Tg WT ND Col1a1-INSR ND Tg Col1a1-INSR HFD Tg Col1a1-INSR Tg Col1a1-INSR Tg
Transcript of Figure S1. Increasing insulin signaling in osteoblasts...
Figure S1. Increasing insulin signaling in osteoblasts weakens glucose intolerance in mice fed a HFD.
(A) Histomorphometric analysis of vertebrae of WT and α1(I)Col-InsR mice fed a ND (n=8), (B) Serum Ctx levels in WT and α1(I)Col-InsR mice fed a ND, (C) Serum osteocalcin levels in WT and α1(I)Col-InsR mice fed a ND (n=8), (D) qPCR analysis of the expression of Glut1, Glut3 and Glut4 in bones of WT and α1(I)Col-InsR fed a ND or HFD (n=5). (E) Western blot analysis of the phosphorylation levels of the INSR and AKT in liver and WAT of WT and α1(I)Col-InsR mice fed a ND or a HFD. All Western blot experiments were repeated at least three times.*P<0.05.
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Figure S2. Compromising insulin signaling in osteoblasts worsens glucose intolerance in mice fed a HFD
(A) Histomorphometric analysis of vertebrae of InsRosb+/- mice fed a ND (n=8), (B) Serum Ctx levels in InsR fl/+ and InsRosb+/- mice fed a ND, (C) Serum osteocalcin levels in InsR fl/+ and InsRosb+/- mice fed a ND (n=8), (D) qPCR analysis of the expression of Glut1, Glut3 and Glut4 in bones of InsR fl/+ and InsRosb+/- mice fed a ND or HFD (n=5). (E) Western blot analysis of the phosphorylation levels of the INSR and AKT in liver and WAT of InsR fl/+ and InsRosb+/- mice fed a ND or a HFD. All Western blot experiments were repeated at least three times. *P<0.05.
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Insr HFD fl/+
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Figure S3. Insulin resistance develops in bone of mice fed a HFD
(A-B) Random glucose (A) and insulin (B) levels of WT mice fed a ND or HFD (n=8). (C-D) GTT (C) and ITT (D) of WT mice fed a ND or HFD (n=7). (E) ELISA analyses of serum levels of leptin, norepinephrine and epinephrine in mice fed a ND or a HFD (n=5). (F-H) ELISA analysis of serum levels of adiponectin in WT (F), α1(I)Col-InsR (G) and InsRosb+/- mice(H) fed a fed a ND or a HFD (n=6). (I-K) qPCR analysis of the expression of Adiponectin in WAT of WT (I), α1(I)Col-InsR (J) and InsRosb+/- mice (K) fed a fed a ND or a HFD (n=5). *P<0.05.
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Figure S4. HFD decreases bone resorption in an insulin dependent manner.
(A-C) microCT analysis of proximal femur of WT and α1(I)Col-InsR mice fed a ND or HFD (n=4). Trabecular bone volume over total tissue volume (A), Cortical bone volume over total tissue volume (B) and mid-shaft cortical bone thickness (C) were measured. *P<0.05.
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Figure S5. Lipotoxicity caused insulin resistance in osteoblasts
(A). Western blot analysis of levels of P-IRS1 at Serine 307 in primary osteoblasts treated with 200µM stearate or vehicle.
IRS1
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