Role of ferroptosis in the process of diabetes-induced endothelial dysfunction

Figure 3 High glucose and interleukin-1β activate the p53-xCT (the substrate-specific subunit of system Xc^-)-glutathione axis in human umbilical vein endothelial cells. A: Human umbilical vein endothelial cells were transiently transfected with p53 small interfering ribonucleic acid or NC small interfering ribonucleic acid and then treated with NG, high glucose (30 mmol/L) and interleukin-1β (10 ng/mL). After treatment for 48 h, the messenger ribonucleic acid levels of p53 and xCT in human umbilical vein endothelial cells with different treatments were determined using real-time polymerase chain reaction analyses. β-actin was used for normalization; B: The protein levels of p53 and xCT were determined using Western blot assays. β-actin was used for normalization; C: The localization of xCT was detected by immunofluorescence. xCT is labeled green, and CD31 is labeled red. The nucleus is labeled blue. Scale bar: 50 μm; and D: The intracellular glutathione/oxidized glutathione (glutathione/oxidized glutathione) ratio was measured with the glutathione and oxidized glutathione assay Kit. ^aP < 0.05 vs the control group. ^bP < 0.01 vs the control group.