Fig. 7: Sesn2 inhibits the ATF4-CHOP-CHAC1 signaling pathway to protect DCs against LPS-induced ferroptosis.

DCs were treated with Sal (20 mM/L) 1 h before LPS stimulation. Cells cultured for 24 h with only LPS were used as the control. A Expressions of ATF4, CHOP, and CHAC1, as detected by Western blotting, were increased under LPS stimulation. *P < 0.05 for the WT-LPS group vs. the control group; ^#P < 0.05 for the Sesn2^−/−-LPS group vs. the control group; ^&P < 0.05 for the Sesn2^−/−-LPS group vs. the WT-LPS group. B Expressions of signaling pathway-related proteins including ATF4, CHOP, CHAC1 and the ferroptosis-related proteins xCT, GPX4, ACSL4, and TFRC in DCs pretreated with Sal before LPS stimulation, were determined by immunoblotting. C–E Levels of GSH, Fe²⁺, and ROS in DCs were measured by detection kits. F Percentages of costimulatory phenotypes expressed on DCs were detected by flow cytometry. G–J The release of IL-12, IL-6, IL-1β, and TNF-α, reflecting the degree of DC maturity and secretion was measured by ELISA. K–L DCs were cocultured with CD4⁺ T cells incubated with Con A to further determine the levels of IFN-γ and the IFN-γ/IL-4 ratio by ELISA. M Flow cytometry results. Data were presented as the mean ± SD from three replicates (n = 3 per group). Statistical significance: *P < 0.05, **P < 0.01 for the WT-LPS + Sal group vs. the WT-LPS group; ^#P < 0.05, ^##P < 0.01 for the Sesn2^−/−-LPS + Sal group vs. the Sesn2^−/−-LPS group; ^&P < 0.05, ^&&P < 0.01 for the Sesn2^−/−-LPS + Sal group vs. the WT-LPS + Sal group.