Fig. 3: Sesn2 inhibits ferroptosis of DCs under LPS stimulation.

A, B Expression of Sesn2 was analyzed by immunoblotting after DCs were stimulated with different doses (40 ng/ml, 200 ng/ml, 1 μg/ml, 5 μg/ml) of LPS for various durations (0, 3, 6, 12, 48 h). β-actin was as the base standard. C–E DCs were isolated from splenocytes of WT mice and Sesn2^−/− mice. The contents of GSH, Fe²⁺, and ROS in DCs were measured by detection kits. F Expressions of xCT, GPX4, ACSL4, TFRC, and Sesn2 were assessed by Western blotting. G Intracellular iron in DCs stained with FerroOrange probes was measured with fluorescence intensity by confocal laser scanning microscopy (×600 and ×1200), scale bar = 25 μm, scale bar = 10 μm. H Lipid peroxidation of DCs was detected by C11-BODIPY after exposure to LPS for 24 h. Red fluorescence represented nonoxidized lipids while green fluorescence was a landmark for oxidation (×600 and ×1200), scale bar = 25 μm, scale bar = 10 μm. I Transmission electron microscopy was applied to observe the morphology of ferroptosis in DCs exposed to LPS, which showed cytoplasmic dilatation and mitochondrial cristae dysfunction (500 nm and 100 nm). Results were shown as the mean ± SD of three replicates (n = 3 per group). Statistical significance: *P < 0.05, **P < 0.01 vs. the WT-control group; ^#P < 0.05, ^##P < 0.01 vs. the Sesn2^−/−-control group; ^&P < 0.05, ^&&P < 0.01 as the Sesn2^−/−-LPS group vs. the WT-LPS group.