Fig. 7: Ferroptosis propagates via transfer of lipid peroxidation reactions between proximal plasma membranes of neighboring cells.
From: Ferroptosis spreads to neighboring cells via plasma membrane contacts
a Scheme of experimental design in (b). Opto-Ctrl or Opto-GPX4Deg and bystander HeLa cells seeded on a supported lipid bilayer or directly glass were exposed to activating illumination. The lipid bilayer is meant to act as a membrane bridge between cells and to facilitate the diffusion of oxidized lipids. Created in BioRender. Garcia, A. (2025) https://BioRender.com/v39k921. b Kinetics of cell death for activated Opto-GPX4Deg and bystander HeLa cells grown or not on a lipid bilayer and treated or not with 5 µM Fer−1. c %AUC for the indicated cell populations. Experiments were performed with three independent biological replicates (n = 3). d Images of C11-Bodipy oxidation and cell death in activated Opto-GPX4Deg HeLa cells and bystander neighboring cells grown on a lipid bilayer. Opto-GPX4Deg, blue; oxidized Bodipy, green; reduced Bodipy, red. Scale bar, 30 µm. e Quantification of oxidized C11-Bodipy over time in cells and in the lipid bilayer (d). f–h Kinetics of cell death for activated Opto-GPX4Deg and bystander HeLa cells treated with DMSO, 100 µM DFO or 100 µM 2,2dipyridyl as indicated. i %AUC for the indicated cell populations. Experiments were performed with three independent biological replicates (n = 3). j Images of C11-Bodipy oxidation (green) in a “donor” GUV stained with DiD (cyan) and containing a photosensitizer lipid (DMMB) before and after activating illumination. Lipid peroxidation (measured as increase in oxidized C11-Bodipy fluorescence, green) is spread to an attached “acceptor” GUV lacking DMMB. Reduced C11-Bodipy shown in red. Controlled illumination in a small membrane region of the donor GUV (yellow box) induced light-activated lipid oxidation in the donor GUV and later on in the acceptor GUV. t = 0 min indicates first image after activation. Membrane regions in which the C11-Bodipy oxidation ratio was calculated are shown in white (acceptor GUV) and pink (donor GUV) boxes. Scale bar, 5 µm. k, l Quantification of the ratio between oxidized and reduced C11-Bodipy in donor and acceptor GUVs over time in absence or presence of 10 mM iron (II) perchlorate. Experiments were performed with three independent biological replicates (n = 3). Statistical analysis by two-sided one-way ANOVA corrected for multiple comparisons using Tukey’s multiple comparison test. Exact p values are shown. All experiments were performed with three independent biological replicates (n = 3). Values are displayed as mean ± SD.
