1. Volume 75, Issue 12, Pages 1297-1307 (June 2009)
Phosphate feeding induces arterial medial calcification in uremic mice: role of serum phosphorus, fibroblast growth factor-23, and osteopontin 
Mohga M. El-Abbadi, Ashwini S. Pai, Elizabeth M. Leaf, Hsueh-Ying Yang, Bryan A. Bartley, Krystle K. Quan, Carly M. Ingalls, Hung Wei Liao, Cecilia M. Giachelli 
Kidney International 
Volume 75, Issue 12, Pages (June 2009)
DOI: /ki
Copyright © 2009 International Society of Nephrology Terms and Conditions

2. Figure 1
Aortic calcium content in severely uremic mice and sham controls under normal and high-phosphate feeding conditions. Dotted lines indicate mean values for each group. (P<0.05, analysis of variance); Sham/NP, n=5; Sham/HP, n=6; SU/NP, n=4; SU/HP, n=7. HP, high phosphate; NP, normal phosphate; SU, severe uremia.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

3. Figure 2
Histological analyses of AMC in uremic mouse arteries. (a) Alizarin red staining of vascular tree whole mount. B-M, Representative micrographs of alizarin red–stained paraffin sections of arteries in the different study groups; (b) abdominal aorta; (c) carotid; (d) iliac; and (e), mesenteric artery in uremic, high phosphate–fed mice (scale bar=100 μm for (b–d), and 50 μm for (e)). Extensive mineralization exclusively in the tunica media was observed in the SU/HP group (f, aortic arch and g, aortic arch; scale bar=30 μm, aortic arch). Early mineralization foci were often observed in the elastic lamella (h, aortic arch) and in the extracellular matrix between lamellae (i, aortic arch) in the MU/HP group (scale bar=30 μm). No arterial calcification was observed in the aorta of mice from the Sham/NP group (j), Sham/HP group (k), SU/NP group (l), or MU/NP group (m) (scale bar=25 μm). L=lumen. HP, high phosphate; Mu, moderate uremia; NP, normal phosphate; SU, severe uremia.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

4. Figure 3
Serum phosphorus, calcium, and calcium × phosphorus in severely uremic mice. Serum phosphorus (a), calcium (b), and calcium × phosphorus (c) linear regression between serum phosphorus and aortic calcium (d) in severely uremic mice and sham controls under normal and high phosphate-feeding conditions. Data represent the mean±s.e.m. Statistically significant differences (P<0.05) were observed between groups with same alphabetical notations; Sham/NP, n=5; Sham/HP, n=6; SU/NP, n=6; SU/HP, n=13.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

5. Figure 4
Aortic calcium content in moderately uremic mice and sham controls under normal and high phosphate–feeding conditions. Dotted lines indicate mean values for each group. (P<0.05, analysis of variance); Sham/NP, n=7; Sham/HP, n=7; MU/NP, n=10; MU/HP, n=10.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

6. Figure 5
Serum phosphorus, calcium, and calcium X phosphorus in moderately uremic mice. Serum phosphorus (a), calcium (b), and calcium × phosphorus (c) linear regression between serum phosphorus and aortic calcium (d) in moderately uremic mice and sham controls under normal and high phosphate–feeding conditions. Data represent the mean±s.e.m. Statistically significant differences (P<0.05) were observed between groups with same alphabetical notations; Sham/NP, n=7; Sham/HP, n=7; SU/NP, n=10; SU/HP, n=10.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

7. Figure 6
Serum FGF-23 and osteopontin levels in moderately uremic mice. Serum FGF-23 (a); correlation between serum FGF-23 and aortic calcium (b); serum osteopontin (OPN) (c); correlation between serum OPN and aortic calcium (d); correlation between serum OPN and FGF-23 (e) levels in moderately uremic mice and sham controls under normal and high phosphate-feeding conditions. Data represent the mean±s.e.m. Statistically significant differences (P<0.05) were observed between groups with same alphabetical notations; Sham/NP, n=7; Sham/HP, n=7; SU/NP, n=6; SU/HP, n=6.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

8. Figure 7
Histochemical staining of mildly calcified and non-calcified arteries. (a–d) are adjacent sections capturing an early mineralization focus in the aorta of a uremic, high phosphate–fed mouse. (a) alizarin red–positive foci of elastin calcification (arrowhead); (b) osteopontin staining (brown) was detected both in SMC surrounding the calcification focus (methyl green counterstained nuclei, arrows) as well as along the calcified elastic lamina (arrowhead); (c) SM22α staining (brown) was absent or greatly reduced in the SMC (methyl green counterstained nuclei) near the calcified focus. (d) Hemotoxylin and eosin (H&E)–stained adjacent section indicates intact SMC in the calcified areas e–h are adjacent aortic sections of an U/NP mouse showing (e), the absence of alizarin red–positive staining; (f) lack of osteopontin staining (brown); (g) abundant SM22α staining (brown) in the medial SMC; (h), normal cellular and matrix morphology by H&E staining; (i) and (j), Runx2/cbfa1-positive nuclear staining (brown) in the aorta of an U/HP mouse (i) or Sham/NP mouse (j). Scale bar 30 μm.
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions

9. Figure 8
Immunochemical staining of a severely calcified artery showing evidence of SMC loss. (a) H&E staining (scale bar=200 μm), boxed area is shown at higher magnification in (b–d); (b) Alizarin red staining (scale bar=30 μm); (c) H&E staining (scale bar=30 μm); (d) SM22α immunostaining (brown) with methyl green nuclear counterstaining (scale bar=30 μm). Note the absence of both SM22α staining as well as cell nuclei in the calcified area (dashed rectangle).
Kidney International  , DOI: ( /ki )
Copyright © 2009 International Society of Nephrology Terms and Conditions