Figure 7.

Histomorphometric characterization and Immunolabeling of Runx-2 and TRAP in WT and 5LOKO mice: (A) Representative images for Runx2 immunostaining and TRAP immunostaining (arrows) of male and female, young (3 months old), and aged (18 months old) 129/SvEv WT and 5LOKO mice. Identification of antigen–antibody reaction was performed using DAB, counterstained with Mayer's hematoxylin. (B,C) Pixels quantification of Runx2 (B) and TRAP (C) was performed by the software Aperio Image Scope v/ 12.3.3 (Leica Biosystems, Buffalo Grove, USA). Images were analyzed with algorithm Positive Pixel Count version 9 and are expressed as positivity (umber of positive pixels/number total). Negative controls were used for calibration and confirmation. Results were presented as the mean and SD for each marker. Different letters indicate significant differences in between young (3 months old) and aged (18 months old) mice for each sex and strain (p < 0.05); symbol * indicates significant differences between WT vs. 5LOKO at the same age.