Fig 2: Deficiency of both GPX4 and vitamin E impairs hematopoietic system homeostasis.a The body weight, b spleen weight, and c total BM cell counts of the Gpx4flox/flox Vav-Cre mice were measured after 3 weeks of a VE-depleted diet. d Left panel: representative flow cytometric plots of the T cell, B cell, and myeloid cell groups in peripheral blood. Right panel: statistical data. e The number of HSPCs in the Gpx4flox/flox Vav-Cre mice after 3 weeks of a VE-depleted diet. N = at least four mice in each group. Data are the mean ± SD. (*P < 0.05, **P < 0.01, ***P < 0.001).

Fig 3: GEN inhibits MI-mediated ferroptosis. (A) Prussian blue staining of iron deposition in rat heart tissues. (B,C) MDA and GSH contents in rats of Sham, MI, MI + DFO, and MI + GEN group. (D) Expression levels of the ferroptosis marker Ptgs2 in rat heart tissues. (E) Grsf1 and GPx4 expression levels in the heart tissues. Data were expressed as mean ± SD (n = 8). *p < 0.05, **p < 0.01 vs. Sham group; # p < 0.05, ## p < 0.01 vs. MI group. Scale bar = 50 µm.

Fig 4: Silencing or inhibition of HO-1 in an appropriate range can reverse Lut-induced ferroptosis in ccRCC by restraining Fe2+ production. (a) Heme levels in 786-O cells treated under different conditions. (b) Levels of intracellular ferrous ion of 786-O cells treated under different conditions. (c) Expression levels of HO-1, GPX4, and SLC7A11 in 786-O cells treated under different conditions as determined by western blot. (d) Semiquantification of band densities in (c). Representative images (e) and semiquantitative analyses (f) of lipid peroxidant in 786-O cells treated under different conditions (magnification, ×200). (g) Interactions of Lut with HO-1 as predicted by molecular docking technology. The data in (a), (b), (d), and (f) are expressed as the mean ± SD. *P < 0.05 vs. the NC group, #P < 0.05 vs. Lut+siNC group, and &P < 0.05 vs. Lut+DMSO group. Lut: luteolin; Znpp: zinc protoporphyrin.

Fig 5: The accumulation of Fe2+ and upregulation of HO-1 are involved in Lut-induced cell death in ccRCC. Levels of intracellular ferrous Fe2+ (a) and GSH (b) of 786-O and OS-RC-2 cells treated with or without Lut for 24 h or 48 h. (c) The LDH release assay showed that Lut-induced cell death of ccRCC was alleviated in the presence of DFP, NAC, and Fer-1 and was exacerbated in the presence of FAC. (d) Expression levels of MMP2, vimentin, E-cadherin, HO-1, KEAP1, NRF2, GPX4, SLC7A11, SLC40A1, DMT1, FTL, and FTH1 in 786-O and OS-RC-2 cells treated with or without Lut for 24 h as determined by western blot analysis. (e) Semiquantification of band densities in (d). The data in (a), (b), (c), and (e) are expressed as the mean ± SD. *P < 0.05 vs. the NC group, #P < 0.05 vs. Lut group. Lut: luteolin; DFP: deferiprone; NAC: acetylcysteine; Fer-1: ferrostatin-1; FAC: ammonium iron (III) citrate.