Fig. 5

Loss of endothelial SETD7 reverses GPX4-mediated lipid peroxidation and oxidative stress. RAECs were cultured in different glucose concentration conditions (Control: 5.5 mM glucose + 16.5 mM mannitol, HG: 22 mM glucose) with or without siScr or siSetd7 treatment. A Representative images showing ROS production. Scale bars, 100 μm. B Representative images of membrane potential detected by JC-10 fluorescence staining. Scale bars, 100 μm. C MDA level, GSH-Px activity, and SOD activity of siScr and siSetd7 RAECs with or without HG stimulation. D The protein expression of GPX4, NOX4, and xCT in siScr and siSetd7 RAECs with or without HG stimulation. E Representative images of immunofluorescence staining of GPX4 in siScr and siSetd7 RAECs with or without HG stimulation. Scale bars, 50 μm. RAECs were transfected with vector (Vector) or Setd7 overexpression plasmid (Setd7^oe). F: Representative images of ROS production and JC-10 fluorescence staining. Scale bars, 100 μm. G MDA level and SOD activity of RAECs transfected with Vector or Setd7^oe. H The protein expression of GPX4 and NOX4 in RAECs transfected with Vector or Setd7^oe. Statistical significances were calculated using (A–C) one-way ANOVA, (D) two-way ANOVA, Tukey’s multiple comparisons tests, (F–H) two tail student’s t test. Data are expressed as the mean ± SEM, statistical analysis revealed a significant difference with **p < 0.01, ***p < 0.001, #p < 0.05, ##p < 0.01, ###p < 0.001, ns = not significant