Figure 1.

Craniofacial skeletal features of END mice and wildtype mice at newborn stage. (A) The skeletal staining of whole mount neonatal END mouse and wild type littermate showing overall changes of skeletal system (top panel), and the confirmation of lack of mRNA by real time PCR (middle panel) and Nell-1 protein by Western Blot (bottom panel) from the calvaria of END newborns. (B) Lateral view of craniofacial skeleton revealed wider coronal sutures (arrows in black), dysplasia of maxillary bones (arrows in yellow) and of auricular bones (arrows in red) in END mouse in comparison to wild type littermate. Scale bar: 1mm. (C) Lateral and Top view of microCT images of neonatal END and wild type skulls demonstrating the difference in cranial bone formation and sutural patency. The red arrows point to auricular bony capsule and the location of the anterior fontanel of calvaria is marked with red asterisk. END images demonstrate widely patent midline sutures and fontanels. Defective mineralization and bone formation is also noted in the END mice. F: frontal bone; M: mandible; P: parietal bone; iP: interparietal bone; sO: supraoccipital bone. Scale bar: 1mm. (D) The red line drawn through the most prominent points of both parietal bones (top image) indicating a cut-plane with labels d1-d4 (bottom image). The d1 represents sagittal suture width, while d2 through d4 are equal 0.5mm interval measuring points of parietal bone thickness starting from d2. Quantitative analysis of the average width and thickness are depicted graphically with significant differences (*P<0.05 and **P<0.01). (E) Depiction of the Head Measurement angulation measurement (left panel). Head angle (°) measured using the angle between two constructed planes, CVT, line representing the long axis of the cervical vertebrae parallel and MPL, mandibular plane line, line representing the mandibular plane. Quantitatative analysis of the head angulation at newborn stage is significantly greater (*P<0.05) in wild type than that of END mice (right graph).