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Assessment of Alizarin Red S Positive Area. a Alizarin Red S (ARS) stained transverse sections of aortic valves of control and pro calcific mice from 4, 16 and 28 weeks. Mean percentage area covered by ARS positive region of the b leaflets, c commissures and d root of aortic valves of control and pro calcific mice from 4, 16 and 28 weeks. Scale bars = 500 µm. N = 2–3. **p < 0.0001, *p < 0.05. Two-way ANOVA with Tukey’s HSD post-hoc multiple comparisons
