Fig 1: IL-38 Suppresses Oxidative Stress in Vascular Calcification. (A) Gene ontology (GO) analysis showing major related molecular function signaling pathways in aortas from WT mice and IL-38 KO mice with VitD3 treatment. (B) Gene set enrichment analysis (GSEA) showing enrichment of genes involved in antioxidant activity. (C) Heatmap of down-regulated genes in antioxidant activity signaling pathway (n = 4). (D) Representative images of DCFH-DA staining of intracellular ROS in hASMCs. (E) Quantification of DCFH-DA staining of intracellular ROS in hASMCs (n = 4). (F) Quantification of malondialdehyde (MDA) of hASMCs (n = 6). (G) Representative images of immunofluorescence staining of 4-HNE (Red), α-SMA (Green) and DAPI (Blue) in hASMCs, scale bar = 50 μm. (H) Quantification of immunofluorescence staining (G) (n = 4). (I) Quantification of superoxide dismutase (SOD) activity of hASMCs (n = 6). (J) Relative mRNA expression of GPX3 from hASMCs (n = 6). (K) Representative Western blot results of GPX3 from hASMCs. (L) Quantification of immunoblots (K) (n = 6). Statistical significance was evaluated via two-way ANOVA followed by the Bonferroni multiple-comparison test (E, F, H, I, J, L). All data were presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001