Fig. 4
From: Hinokitiol-iron complex is a ferroptosis inducer to inhibit triple-negative breast tumor growth
Fe(hino)3 -induced ferroptosis is iron-dependent. A–C The content of GSSG and GSH (A), the expression of HO-1, GPX4, xCT, and SOD2 (B), and the enzyme activity of GPX4 (C) in MDA-MB-231 cells treated with Fe(hino)3 (5 µM) for 24 h. D The expression of Ferritin, TfR1, NRF2, and GPX4 in MDA-MB-231 cells treated with 5 µM Fe(hino)3 alone or with Trolox (200 µM) or NAC (5 mM) together for 24 h. E Total iron content measured by ferrozine assays in MDA-MB-231 cells treated with Hino (15 µM), ferric ammonia citrate (FAC) (5 µM), and Fe(hino)3 (5 µM) for 24 h. F Cellular nonheme iron levels were detected using DAB-enhanced Perl’s staining. G Upper panel: Ferritin (FTH subunit) expression in MDA-MB-231 cells treated with different concentrations of FAC or Fe(hino)3 for 24 h, detected by immunoblotting. Lower panel: the iron level in ferritin was detected by Perl’s stain. H–I Cell viability (H) and protein expression (I) of MDA-MB-231 cells after 24-h incubation with Fe(hino)3 (5 µM) or/and DFO (50 µM) or DFP (50 µM). J The lipid ROS in MDA-MB-231 cells treated with Fe(hino)3 (2 µM) alone or with FAC (5 µM) together for 24 h, detected by flow cytometry and indicated by BODIPY-C11. K Cell viability after cotreatment with Fe(hino)3 (2 µM) and FAC for 24 h. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001