Fig. 2

CSE triggered ferroptosis and upregulated the expression of plasmalogen biosynthesis in human alveolar epithelial cells. Human type II alveolar epithelial A549 cells were exposed to 0.1%, 0.5%, 2%, and 5% CSE for 24 h. (A) Cell viability was detected by CCK-8 assay. (B) LDH release was detected by LDH activity assays. (C) The concentration of IL-33 and IL-1α in the cellular supernatant was determined by ELISA assay. (D) The levels of total ROS were determined using a 2’, 7’-dichlorofluorescein diacetate (DCFH-DA) kit. (E-G) Quantification of the ferroptosis-related markers MDA, GSH, and GPX4. (H) The protein levels of GPX4, FAR-1, AGPS, and GNPAT were detected by western blot analysis. (I) The mRNA expression of FAR-1, AGPS, and GNPAT was measured via quantitative real-time PCR. Data are presented as the mean ± SD of three replicates and analyzed using one-way analysis of variance (ANOVA), followed by Dunnett’s test. ^nsp > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001, compared with blank. The gels were cropped reasonably