Extended Data Fig. 2: Inactivation of lysosomal iron inhibits ferroptosis (Part 2).
From: Activation of lysosomal iron triggers ferroptosis in cancer
a, Chemical syntheses of metcLip-1 and alcLip-1. b, Fluorescence imaging of labelled cLip-1 (10 µM, 2 h), metcLip-1 (10 µM, 2 h) and alcLip-1 (10 µM, 2 h). Scale bar, 10 μm. c, Cyclic voltammetry of a FeCl3 solution (reduction potentials = pink arrow). Data recorded in the presence of metcLip-1 or alcLip-1. d, Bodipy-C11 581/591 flow cytometry of cells treated with metcLip-1 (10 µM), alcLip-1 (10 µM) and RSL3 (100 nM) for 1 h. Representative of n = 3 independent experiments for PDAC053T and n = 1 for HT-1080. e, FENIX assay of cLip-1, metcLip-1 and alcLip-1. f, Molecular structure of cDFO and fluorescence imaging of labelled cDFO (100 µM, 15 min). Scale bar, 10 μm. n = 3 independent experiments. Data are mean ± s.d. g, Fluorescence emission spectra of RPE at pH 5 in the presence of FeCl3 and Lip-1. h, RPE flow cytometry of cells treated for 30 min and then cotreated with the probe for 30 min. HCQ (100 µM). Representative of n = 3 independent experiments. i, HMRhoNox-M flow cytometry of cells treated for 30 min and then cotreated with the probe for 30 min. HCQ (100 µM). Representative of n = 3 independent experiments. j, Bodipy-C11 581/591 flow cytometry of cells treated with RSL3 (200 nM for PDAC053T, 500 nM for 4T1, 1 h) and Baf-A1 or HCQ (10 µM) used 2 h prior. Representative of n = 3 independent experiments. k, Fluorescence imaging of Bodipy 665/676 in cells treated with RSL3 (1 µM, 1 h). Scale bar, 10 μm. n = 3 independent experiments. Two-sided unpaired t-test. Data are mean ± s.d. Concentrations used unless stated otherwise: Lip-1 (10 µM), Baf-A1 (75 nM).
