JCS00772 Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Transgenic JunB expression and normal endochondral ossification during embryogenesis. (A) In situ detection of JunB mRNA in embryonal bone sections (E18,5) of control and junB^-/- Ubi-junB littermates. Sections were counterstained with eosin and haematoxylin and images were taken with 10-fold magnification. Normal embryonal chondrogenesis was demonstrated by in situ detection of type-II collagen (col-II) positive proliferative and prehypertrophic chondrocytes in femoral sections (B) and normal bone formation was shown by alizarin red staining of femur, rib and tail sections from 18,5-day old control and mutant embryos (C).

• Supplemental Figure 2 -

  Fig. S2. Reduced bone mass in calvaria of junB^-/- Ubi-junB mice. 10 mm sections of 4% PFA fixed, decalcified and paraffin embedded calvaria from 16-week old control (A, B) and mutant (C, D) skulls were analysed by eosin and haematoxylin staining. Reduced bone mass demonstrates that osteoporosis is not restricted to trabecular and cortical bone formed by endochondral ossification, but is also present in bones formed by intramembraneous ossification.

• Supplemental Figure 3 -

  Fig. S3. Impaired angiogenesis in long bones of junB^-/- Ubi-junB mice. (A) Eosin and haematoxylin staining (HE) of tibiae sections from 4-week old littermates revealed reduced number of blood vessels filled with erythrocytes in junB^-/- Ubi-junB animals (top panel, magnification 10-fold). Reduction was still present in vertebrae of the tail from 16-week old mutant animals (middle panel, magnification 20-fold) and was correlated with reduced signal for the endothelial marker CD31 measured by immunofluoreszence (bottom panel). Sections were counterstained with Hoechst dye (B) Corresponding to the functional link between angiogenesis and apoptotis of hypertrophic chondrocytes, we observed by in situ hybridization an increase in type-X collagen (col-X) positive chondrocytes in tibiae sections of 16-week old  junB^-/- Ubi-junB compared to control littermates. Sections were counterstained with eosin and haematoxylin and images were taken with 40-fold magnification.