Fig. 1.

Loss of ABCC6 predisposes skeletal muscle to nanohydroxyapatite deposition at 7DPI. a WT (Abcc6^+/+), heterozygous (Abcc6^+/−), and homozygous (Abcc6^−/−) mice were assessed for calcification at the site of skeletal muscle injury by radiographic analysis and subsequent STiCSS quantification at 7 DPI. See Table 2 for detailed analysis of the genotypes and N. ***p < 0.001; ****p < 0.0001. Statistical analysis between groups was performed using a non-parametric Mann–Whitney test. b Representative 3D µCT reconstructions and histological analysis of skeletal muscle calcification within the injured gastrocnemius and soleus muscles at 7 DPI. Scale bar represents 100 µm. n ≥ 3 mice per genotype. Positive Von Kossa staining, noted by the black deposits, indicates calcium deposition within the damaged skeletal muscle. c Energy dispersive X-ray (EDS) analysis of dystrophic calcification nodules within damaged ABCC6-deficient skeletal muscle at 14 DPI. Topographic mapping demonstrated marked co-localization of calcium and phosphate with an average calcium/phosphate atomic ratio of 1.67 ± 0.2, indicative of hydroxyapatite. Analysis was conducted following random sampling of 5 distinct spots per tissue section. Scale bar represents 200 µM, thereby indicating nanohydroxyapatite