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. Author manuscript; available in PMC: 2016 May 1.
Published in final edited form as: Exp Cell Res. 2015 Feb 20;333(2):289–302. doi: 10.1016/j.yexcr.2015.02.009

Figure 2.

Figure 2

High-magnification immunohistochemistry of decalcified Col1(2.3)/GFP/Rs1 (central area) and control (whole area) calvariae from 9-week-old mice. (A) Osterix immunohistochemistry on formalin-fixed samples, demonstrating significant nuclear staining (brown; red arrowheads) by cells throughout the Col1(2.3)/GFP/Rs1 bony lesion mostly between the trabeculi. (B) Osteocalcin immunohistochemistry on formalin-fixed samples, demonstrating that cytoplasmic (brown; blue arrowhead) and extracellular (brown; red arrowhead) osteocalcin are present in the bone lesions, confirming the presence of mature osteoblasts. (C) FLAG immunohistochemistry on ethanol-fixed samples showed cytoplasmic staining of Rs1 expression only in Col1(2.3)/GFP/Rs1 calvarial sections (brown; red arrowhead). (D) CD31 immunohistochemistry on formalin-fixed samples, demonstrating diffuse positivity of blood vessels and sinusoids in the mutant clavaria (brown; red arrowheads). (Scale bar 100μm) tb, tabeculi