Fig. 1.

Vascular nesfatin-1 expression is upregulated under calcifying conditions. (A, B) Western blot analysis of nesfatin-1 protein levels in VSMCs with high Pi (2.6 mM). (C) RT-PCR analysis of nesfatin-1 mRNA levels in VSMCs with high Pi (2.6 mM). (D, E) Western blot analysis of nesfatin-1 protein levels in VSMCs with β-GP (5 mM) and Ca²⁺ (2.5 mM). (F) RT-PCR analysis of nesfatin-1 mRNA levels in VSMCs with β-GP (5 mM) and Ca²⁺ (2.5 mM). (G) Western blot analysis of nesfatin-1 protein levels in isolated aortas with or without high Pi (3.8 mM) for 7 days. (H) RT-PCR analysis of nesfatin-1 mRNA levels in isolated aortas with or without high Pi (3.8 mM) for 7 days. (I) Immunofluorescence analysis of nesfatin-1 in isolated aortas with or without high Pi (3.8 mM) for 7 days. Scale bar, 50 μm. (J) Western blot analysis of nesfatin-1 protein levels in aortas from VD3-induced mice. (K) RT-PCR analysis of nesfatin-1 mRNA levels in from VD3-induced mice. (L) Immunofluorescence analysis of nesfatin-1 in isolated aortas from VD3-induced mice. Scale bar, 50 μm. (M) Serum levels of nesfatin-1 in different VC groups. (N) The positive association of serum nesfatin-1 levels with the coronary artery calcium scores by a Spearman correlation analysis. Differences between groups were assessed with ANOVA followed by Bonferroni post-hoc test (B-C, E-F, M). The P-value was calculated by unpaired two-tailed Student’s t-test (G-H, J-K). The statistical significance of correlations was assessed by spearman’s correlation coefficient analysis (N). *P < 0.05, **P < 0.01, ***P < 0.001 versus the indicated group. n = 6