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. Author manuscript; available in PMC: 2013 Jan 1.
Published in final edited form as: Nat Med. 2012 Jul;18(7):1095–1101. doi: 10.1038/nm.2793

Figure 2. Suppressed osteoblast maturation in Igf1r−/− (Osx-Cre; Igf1rfl/fl) mice.

Figure 2

(a) Immunohistochemical analysis of Runx2, Osterix, and Osteocalcin performed on trabecular bone sections from distal femora of 3-month-old female Igf1r−/− (Osx-Cre; Igf1rfl/fl) mouse and wild type littermate mouse Igf1r+/+ (Osx-Cre). Osx-GFP expressing cells observed by direct fluorescence microscopy appear green, immunofluorencent staining for osteocalcin visualize red (far right panel). Scale bar: 100 µm. (b–d) Numbers of Runx2, Osterix, and Osteocalcin positive cells on bone surface, measured as cells per millimeter of perimeter in sections. n = 5. *p < 0.05. (e,f) Light micrographs of tartrate-resistant acid phosphatase (TRAP)-staining performed on trabecular bone sections from distal femora of mice. Number of osteoclasts per tissue area (N.Oc/T,Ar) was measured. Data represent the mean ± SEM. n = 10. *p < 0.05. (Scale bar: 100 µm) (g) CFU-F and CFU-Ob assays from harvested bone marrow of the mice as indicated. Representative images of CFU-Fs stained with crystal violet (top panels). Representative images of CFU-Obs stained with Alizarin Red (bottom panels). (h,i) Quantifications of the CFU-F and CFU-Ob assays. Data represent the mean ± SEM. of triplicate cultures of bone marrow nucleated cells pooled from five individual mice. *p < 0.05.