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. 2012 Aug 21;7(8):e43372. doi: 10.1371/journal.pone.0043372

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© 2012 Iwasaki et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A, Runx2 expression (arrowheads) in the posterior longitudinal ligament (arrows). In situ hybridization of Runx2 in wild-type (WT) mouse vertebrae at birth (left). LacZ staining in WT mouse vertebrae at birth (middle). Immunohistochemistry of Runx2 in WT mouse vertebrae at embryonic day 16.5 (right). B, Radiographic assessment of the development of calcification of the ligament in an Enpp1^ttw/ttw mouse at 4 and 8 weeks of age. Note an appearance of calcification at 8weeks (arrowheads) C, Histological assessment of the cruciform ligament (arrowheads) at the atlanto-occipital area in an Enpp1^ttw/ttw mouse at 4 and 8 weeks of age. D, Immunohistochemical staining of Runx2 at the posterior longitudinal ligament in an Enpp1^ttw/ttw mouse at 4 weeks of age. Note that Runx2 was expressed in an area corresponding to the prospective calcification.