Adenine-treated rats develop renal dysfunction prior to media calcifications and increased in vivo stiffness being present. Serum creatinine (A) and total aortic calcium (B) are shown for each timepoint (2 and 4 weeks). Abdominal aPWV (C) and distensibility (D), measured by ultrasound, are shown for each timepoint (2 and 4 weeks). Number of animals per group (A–D): control 2 and 4 weeks: n = 7; adenine 2 and 4 weeks: n = 7. Ordinary one-way ANOVA with Bonferroni multiple comparison test (A–D): P > .05: not significant, not shown; *P < .05; **P < .01;****P < .0001. Data in bar graphs are represented as mean ± SEM. Formalin fixed, paraffin-embedded thoracic aorta cross sections (50× magnification) are used to visualize mineralization by the Von Kossa method (E). Calcium phosphate depositions (Von Kossa positive) are depicted as black precipitation. Cross sections are separated per condition (Ctrl, CKD) and per timepoint (2 and 4 weeks). Number of animals per group (E): control 2 and 4 weeks: n = 7; adenine 2 and 4 weeks: n = 7. Scale bar: 500 μm.