Figure 4

Figure 4

NARFL Gene Deletion Leads to Endothelial Cell Dysfunction. A) Cell Rox kit was used to detect the oxidative stress level of endothelial cells. The darker the color of red fluorescence, the higher the level of cellular oxidative stress. B) Quantitative results of cellular oxidative stress were detected by CellRox kit; the results showed that NARFL deletion led to an increase in oxidative stress level of endothelial cells, and ferroptosis inhibitor Ferrostatin‐1 could partially alleviate oxidative stress injury caused by NARFL gene deletion. C) The expression levels of endothelial cell marker CD31 and fibroblast marker α‐SMA in HPMEC and NARFL^−/− HPMEC were detected by immunofluorescence. The fluorescence intensity directly correlates with protein expression levels, and the result showed that deletion of the NARFL gene led to lower CD31 (red) expression and up‐regulation of α‐SMA (green). D‐F) HPMEC, NARFL^−/− HPMEC, and HPMEC transfected NARFL point mutant (c.482 G> T) mitochondrial respiratory function curve, black represents wild type, green represents NARFL (c.482 G>T) point mutant, with red representing HPMEC with NARFL deletion mutations. The results showed that mitochondrial respiratory function decreased with NARFL (c.482 G>T) point mutant, but there was little difference compared with wild‐type cells, while NARFL deletion mutation resulted in significant decrease of mitochondrial function. 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.