Fig. 4

MICU1 deficiency aggravates the apoptosis, inflammatory response and nitrification stress of CMECs treated with HGHF. (A) KEGG pathway enrichment analysis based on these 496 overlapping DEGs. The top 10 enriched pathways ranked by the number of DEGs in the pathways are shown. (B) Heatmap showing the expression levels of DEGs belonging to apoptosis and the NF-κB signaling pathway in RNA-Seq. The color scale depicts the range of log2-fold changes in gene expression. (C) Six selected DEGs in this pathway were validated using qRT-PCR (n = 6 per group). (D) Flow cytometry of apoptosis using annexin V and propidium iodide (PI) staining in CMECs treated as indicated (n = 6 per group). (E) Changes in the expression of NLRP3, NF-κB, Caspase 1, IL-1β,TNF-α and Cleaved caspase 1as assessed by Western blotting (n = 4 per group). (F) Western blotting of eNOS, p-eNOS, iNOS and β-actin in CMECs (n = 4 per group). (G) Analysis of 3-NT Elisa results of CMECs (n = 6 per group). (H) FITC-dextran clearance was measured to assess changes in endothelial permeability (n = 6 per group). *p < 0.05, **p < 0.01