Extended Data Fig. 6: Identification of ferroptosis-resistant cell states perturbed by 2,4-DIH.
From: High-throughput profiling of chemical-induced gene expression across 93,644 perturbations
a-c, Cell viability of HK-2 cells after treatment with ERA (a), cisplatin (b) and 2,4-DIH (c) for 48 h (n = 3 biological replicates). d-f, HK-2 cells were treated with 2,4-DIH (250 μM) and ERA (panel d: 10 μM, panel e: 30 μM, panel f: 15 μM) for 24 h. Subsequently, the cellular ROS was tested by flow cytometry (d). HK-2 cells were labeled with FerroOrange fluorescent probes to measure intracellular Fe2+ concentrations using flow cytometry (e). Lipid peroxidation in HK-2 cells was detected using flow cytometry (n = 3 biological replicates, one-way ANOVA) (f). g, Representative photomicrographs of transmission electron microscopy in HK-2 cells. HK-2 cells were treated with 2,4-DIH (250 μM) and ERA (20 μM) for 24 h. Blue arrows indicate normal mitochondria. Red arrows indicate abnormal mitochondrial morphology typical of ferroptosis, including shrunken mitochondria, increased density and rupture of membranous structure, and decreased mitochondrial cristae. Yellow arrows indicate mildly abnormal mitochondria. Data shown represent mean ± SD. 2,4-DIH, 2,4-dihydroxybenzaldehyde. ERA, erastin. DCFH-DA, 2,7-dichlorofluorescein diacetate. C11-BODIPY581/591, 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3- propionic acid.
