Fig. 2

Application of the dual-vector dCasRx translation enhancement tool in a CaOx crystal-induced cell injury model. A Volcano plot shows differential expression genes (DEGs) between CaOx crystal-induced cell injury models and negative controls. B The protein expression of FTH1, GPX4, ASCL4, and CD71 (also called TFRC) in models were determined by 4D-LFQ proteomic analysis. C After constructing the dual-vector CRISPR-dCasRx-eIF4GI tool targeting FTH1 (DV-FTH1), the mRNA expression level of FTH1 were detected using qRT–PCR analysis. D, E WB analysis results show the protein expression levels of FTH1, CD71 GPX4, and ASCL4 between CaOx + Neg. and CaOx + DV-FTH1, and the bar graph shows the relative protein levels. F The cell viability was detected using CCK8 assay kit. G Several indicators of anti-oxidative status, including T-AOC content, SOD content and CAT content, were measured. H Two indicators of ferroptosis, including MDA content and Fe²⁺ content, were measured. I The level of cellular ROS was observed and images were taken under dark field (magnification ×400). Brighter green indicates higher levels of cellular ROS. J The level of cellular Fe²⁺ was observed and images were taken under dark field (magnification ×400). Brighter red indicates higher levels of cellular Fe²⁺. K The degree of lipid peroxidation was observed and images were taken under dark field (magnification ×400). Higher ratio of green to red indicates higher degree of lipid peroxidation. L The mitochondrial morphology was observed using transmission electron microscopy (scale bars, 2 μm or 1 μm). Data are presented as the means ± SEM from three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and ns, no significant difference, represents P > 0.05