Fig. 6
From: CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD
Experimental validation of CSE-induced apoptosis and ROS generation via SIRT4-mediated GNPAT acetylation. A549 cells were classified into four groups, including blank + vector, 5% CSE + vector, 5% CSE + SIRT4 and 5% CSE + SIRT4 + GNPAT. (A-B) The expression levels of SIRT4 and GNPAT at mRNA and protein were determined in the above A549 cells. (C) Immunofluorescence staining of GNPAT was shown. (D) GNPAT acetylation level was detected by immunoprecipitation assay. (E) Cell viability was detected by CCK-8 assay. (F) The LDH release was detected by LDH activity assays. (G) Cell apoptosis was determined by flow cytometry assay. (H) The levels of total ROS were determined using a 2’, 7’-dichlorofluorescein diacetate (DCFH-DA) kit. Data are presented as the mean ± SD of three replicates and analyzed using ANOVA, followed by Tukey’s post-hoc test. **p < 0.01, ***p < 0.001, compared with blank + vector; #p < 0.05, ###p < 0.001, compared with 5% CSE + vector; $$$p < 0.001, compared with 5% CSE + SIRT4. The gels were cropped reasonably