Fig. 5: LCN2 mediates ferroptosis resistance in ILC3s via the ATF4-xCT/GPX4 axis.
From: GPX4 restricts ferroptosis of NKp46+ILC3s to control intestinal inflammation
MNK3 cells were transfected with si-con / si-Lcn2, or plasmids encoding LCN2 (OELCN2) / empty vector (OEEV) and treated with RSL3 (2-8 μM), erastin (1–4 μM) or vehicle (DMSO) for 16 h. Statistical results of a, c the percentage of annexin V and 7-AAD double-negative population and b, d lipid ROS production in the indicated MNK3 cells. (n = 4) e ATF4, xCT, and GPX4 expression in MNK3 cells following transfection with si-con or si-Lcn2 and treatment with erastin for 16 h. (n = 4) f After transfecting MNK3 cells with si-con or si-Lcn2, the MNK3 cells were subjected to erastin (4 μM) treatment for 12 h in the presence or absence of GSH (0.5 mM) or NAC (0.5 mM), and subsequently, lipid ROS production was assayed. (n = 4) g RORγt, IL-22, and IL-17A expression in MNK3 cells following transfection with si-con or si-Lcn2 and treatment with erastin for 16 h. (n = 3) Statistical results of MFI for h p-p38-Thr180/Tyr182 and i p-ERK1/2 (Thr204/Thr187) expression in the indicated MNK3 cells following treatment with erastin. (n = 3) j ATF4, xCT, and GPX4 expression in MNK3 cells following transfection with plasmids OELCN2 or OEEV and treatment with erastin for 16 h. (n = 4) k RORγt, IL-22, and IL-17A expression was assessed in indicated MNK3 cells treated with erastin for 16 h. (n = 3) Statistical results of MFI for l p-p38-Thr180/Tyr182 and m p-ERK1/2 (Thr204/Thr187) expression in indicated MNK3 cells after treatment with erastin. (n = 3) n, o After transfecting MNK3 cells with plasmids OELCN2 or OEEV, the cells were exposed to erastin for 16 h in the presence or absence of the p38 inhibitor SB203580. n RORγt, IL-22, and IL-17A expression in indicated MNK3 cells. (n = 3) o ATF4, xCT, and GPX4 expression in indicated MNK3 cells. (n = 3) p Statistical results of MFI for p-p38-Thr180/Tyr182 expression in ILC3s from LPMCs of Lcn2fl/fl and RorccreLcn2fl/fl mice post C.R infection. (n = 4) Data are presented as the mean ± SEM or median, and statistical significance was determined using two-sided unpaired t-test (a–p). *P < 0.05; **P < 0.01; ***P < 0.001.
