Fig. 3. Sirtuin 3 (Sirt3) was required in the inhibitory effect of soluble epoxide hydrolase (sEH) deletion on calcium deposition in vascular smooth muscle cells (VSMCs).

A Heatmap of all differential expression genes (DEGs) between Ephx2^−/− (EP) and wild-type (WP) VSMCs with high phosphate (Pi) treatment. B Volcano plots displaying the fold change (log2) of all genes analyzed. C Enrichment plot of Central Carbon Metabolism in Cancer gene set in gene set enrichment analysis (GSEA). D Venn diagram displaying DEGs in the GSEA gene sets. E, F Western blot analysis (A) and its quantitative analysis of Sirt3 (B) between Ephx2^−/− and wild-type (WT) VSMCs treated with high phosphate (Pi). G, H Alizarin S red staining (G) of VSMCs with scramble small interfering RNA transfection (siRNA) or Sirt3 siRNA treatment. The related staining OD values (562 nm) were shown (H). I Quantification of calcium contents of VSMCs. The calcium contents of each group were normalized to the related protein concentrations. J–L Western blot analysis and its quantitative analysis of alpha smooth muscle actin (α-SMA) (K) and bone morphogenetic protein-2 (BMP2) (L) protein expression levels in VSMCs with scramble siRNA or Sirt3 siRNA treatment, respectively, under high Pi stimulation. Data are presented as mean ± SD. *P < 0.05; ^#P < 0.01; ^$P < 0.001; ^&P < 0.0001.