Figure 5

NARFL Gene Deletion Leads to Endothelial Cell Dysfunction with Abnormal Ferroptosis Pathway and Inability to Interact with CIA System‐Related Proteins in Endothelial Cells. A,B) Tube formation experiment compared HPMEC, NARFL^−/− HPMEC, NARFL^−/− HPMEC treated with Ferrostatin‐1 inhibitor of iron ions, and NARFL^−/−HPMEC transfected with CYP2J2 plasmid. The results showed that NARFL deletion would lead to difficulties in tubular formation. However, Ferrostatin‐1 and the overexpression of the CYP2J2 plasmid can partially alleviate the tubule formation difficulty. C,E) Western blot results of HPMEC wild‐type cells, NARFL^−/− HPMEC cells, and NARFL^−/− HPMEC cells transfected with the overexpression of the NARFL plasmid. The results showed that lower NARFL expression led to lower expression of GPX4, SLC7A11 and Ferritin. The expression of TFR1 and IRP1 was up‐regulated, whereas CYP2J2 was barely expressed in NARFL^−/− HPMEC; transfection with an overexpression NARFL plasmid in NARFL^−/− HPMEC partially compensated for these changes. D, Result of co‐immunoprecipitation; the Co‐IP result shows the NARFL mutant could not interact with MMS19, CIAO1 and MIP18. Statistical significance was assessed using one‐way ANOVA with Tukey's multiple comparison correction. ns, not significant; *p < 0.05; **p < 0.01; ***p < 0.001.