Extended Data Fig. 6: Fento-1 induces the oxidation of phospholipids and cell death by activating lysosomal iron (Part 2).
From: Activation of lysosomal iron triggers ferroptosis in cancer
a, CellTiterGlo luminescence of cells treated with Fento-1 (10 µM, 6 h) and inhibitors used 2 h prior. n = 5 independent experiments. b, Resorufin fluorescence of cells treated with Fento-1 (10 µM, 6 h) and inhibitors used 2 h prior. Vehicle, Lip-1: n = 8 independent experiments, clip-1 and Z-VAD-FMK: n = 4 independent experiments, Fer-1, Def and Toc: n = 6 independent experiments, DFO, DFX and Vitamin K3: n = 7 independent experiments. c, d, CellTiterGlo luminescence of cells treated with Fento-1 (10 µM, 6 h) and inhibitors used 2 h prior. n = 7 independent experiments. e, Incucyte cell viability measurement. Representative of n = 3 independent experiments. Data are mean ± s.e.m. f, Viability of cells treated with Fento-1 (10 μM, 24 h) or Lip-1 (1 μM, 24 h) or Fento-1 (10 μM, 24 h) and Lip-1 (1 μM, 30 min prior). 1-way ANOVA. n = 6 technical replicates. g, Fluorescence imaging of Fento-1. Fento-1 (1 μM, 1 h) and Toc (100 μM, 2 h prior). Scale bar, 10 μm. n = 3 independent experiments. Two-sided unpaired t-test. Data are mean ± s.d. h, Viability of cells treated for 48 h. n = 3 independent experiments. Data are mean ± s.d. i, Left: Cell death in knock-down cells measured by Annexin-V/ Sytox blue in cells treated with Fento-1 for 6 h. n = 3 independent experiments. 2-way ANOVA. Data are mean ± s.d. Right: Western blot of proteins in knock-down cells. γ-tubulin is a sample processing control. j, Bodipy-C11 581/591 flow cytometry of knock-down cells treated for 6 h. Box plots: interquartile range, centre lines = medians and whiskers = the minimum and maximum values. a-d, Two-sided Mann–Whitney test compared to vehicle.
