Fig 3. Dietary magnesium supplementation attenuates soft tissue calcification in D2, Ahsg-/- mice.

a-d. Three-week-old mice were fed for eight weeks normal chow containing 0.2% magnesium, or a high magnesium diet containing 1% magnesium. Computed tomography of upper torso shows sparse calcified lesions in mice at the start of the feeding experiment (a,c), massive calcification after eight weeks on normal magnesium chow (b), and strongly attenuated calcification after eight weeks on high magnesium chow (d). e, Calcified tissue volumes determined at age 11 weeks by CT and segmentation. f, Tissue calcium content was measured following chemical extraction of brown adipose tissue (BAT), kidney, lung and heart. g-n show von Kossa stained cryosections of g,k, BAT, h,l, kidney, i,m, lung, and j,n, heart. Scale bar indicates 200 μm. Mice on normal chow had large calcified lesions in their BAT, occasional lesions in kidney and lung, and fibrosing calcified lesions in myocard (b,f,g-j). In contrast, mice fed with high magnesium developed sparse calcified lesions in the same organs (d,f,k-n). o-r, Unlike mice with the calcification resistant genetic background B6, mice with the calcification-prone genetic background D2 failed to induce Trpm6 mRNA expression in the kidney, which resulted in the functional hypomagnesemia described in Fig 1H. Student t-test (e,f) and one-way ANOVA with Tukey multiple comparison test (o-r) for statistical significance, **p<0.01, ***p<0.001.