Figure 3

Characterization of osteoblast organization in femoral sections from Mirc24 cluster-deficient mice. Osteoblast distribution in plane matched sections of femora from newborn male Col2a1-Cre (Cre) and Col2a1-Cre-Mirc24^tm1M/Y (hKO) mice was analyzed by immunodetection of (a) osteopontin and (b) osterix. (c) In addition, alkaline phosphatase (ALP) activity was detected by histological staining. (d,e) To determine osteoblast numbers, paraffin sections were stained with Giemsa and Toluidine Blue. Brightness was adjusted using Adobe Photoshop Elements 2020 (Version 18.0, https://www.adobe.com/products/photoshop-elements.html) for visualization (d,e). At least three animals per genotype were analyzed and representative images are shown. Additional images are shown in Supplementary Figs. S3 and S4 online. (f,g) For quantification, the number of Giemsa⁺ and Toluidine Blue⁺ osteoblasts per mm trabecular bone were determined. Individual and mean values and standard deviations are shown (graph). The figure was created using Adobe Illustrator CS6 (Version 16.0.3, https://adobe.com/products/illustrator).