Fig. 4.

The relationship between Cal and CFA. (A) Development of Cal shown by Alizarin Red staining. Cal first appears at week 25 and rapidly enlarges over the next 5 weeks. (B) Development of Cal shown by Von Kossa staining, with results consistent with those observed in Alizarin Red staining. (C) Magnified images of the Cal site. The matrix and nuclei at the site of Cal are random. (D) Masson staining and statistical analysis of Col fiber orientation distribution, showing that CFA at the site of Cal is random (n = 100). (E) Alizarin Red, Von Kossa, Masson, Sirius Red, and Col I IHC images of the vascular wall at week 30, showing marked colocalization between Cal and Col I. The same mouse was used for each time point and IHC, with a total of 5 mice used.