Fig. 5
MICU1 knockout aggravates endothelial dysfunction by promoting nitrification stress mediated by mitochondrial calcium uptake in endothelial cells. (A) Quantitative determination of Ca2+ content in mitochondria of CMECs from the indicated groups (n = 6 per group). (B) Mitochondrial membrane potentials were analyzed using JC-1 staining in CMECs treated as indicated. Flow cytometry results are presented (n = 6 per group). (C). Representative Western blotting of eNOS, p-eNOS, iNOS and β-actin in CMECs (n = 4 per group). (D) Analysis of 3-NT Elisa results of CMECs (n = 6 per group). (E) Flow cytometry of apoptosis using annexin V and PI staining in CMECs treated as indicated (n = 6 per group). (F) Changes in the expression of NLRP3, NF-κB, Caspase 1, IL-1β, TNF-α and Cleaved caspase 1 assessed using Western blotting (n = 4 per group). (G) FITC-dextran clearance was measured to assess changes in endothelial permeability (n = 6 per group). **p < 0.01