Fig. 5

Interference with GNPAT can alleviate CSE-induced ferroptosis in human alveolar epithelial cells. A549 cells were transfected with sh-GNPAT or sh-NC for 48 h, followed by 5% CSE treatment. (A-B) The expression levels of GNPAT mRNA and protein were determined in the above A549 cells. (C) Immunofluorescence staining of GNPAT was shown. (D) Cell viability was detected by CCK-8 assay. (E) The LDH release was detected by LDH activity assays. (F) The levels of total ROS were determined using a 2’, 7’-dichlorofluorescein diacetate (DCFH-DA) kit. (G) Cell apoptosis was determined by flow cytometry assay. (H) Mitochondrial morphology associated with ferroptosis was determined by transmission electron microscopy. (I) Detection of lipid oxidation by the BODIPY 581/591 C11 probe method. (J) Content detection of the ferroptosis-related markers MDA, GSH, and GPX4 by ELISA. A549 cells were exposed to 0.1%, 0.5%, 2%, and 5% CSE for 24 h, followed by quantification of SIRT4 expression via western blot analysis (K) and GNPAT acetylation level by immunoprecipitation (L). Data are presented as the mean ± SD of three replicates and analyzed using Student’s t-test. ***p < 0.001, compared with sh-NC. The gels were cropped reasonably