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. 2016 Mar 7;4:15. doi: 10.3389/fped.2016.00015

Figure 3.

Figure 3

Comparisons of bone integrity between controls and IH rats. (A) First column: overview from which a–f panels are derived. Immunohistochemical staining using Runx2 antibody shows decreased expression of Runx2 protein (brown color) in the femur, harvested from IH-treated animals (panel d), over control (panel a). Panels b and e contrast the differences in the length of growth plate in the tibia (white scale bar = 500 μm). Panels c and f show cell density of the proliferating and hypertrophic zone of the growth plate (black scale bar = 100 μm). (B) Masson trichrome staining visualizes the zones of proliferation and hypertrophy of growth plate areas. No significant difference in cell counts appears between control and IH-treated bones. Gray scale bars = 300 μm. (C) Western blot assays demonstrate a decreased cellular expression of Runx2, Collagen-1 (COL1), and ZIP8 transporter proteins with respect to β-actin. (D) Zinc content in the tibia (whole bone extracts, n = 3) declined significantly after IH treatment.