Extended Data Fig. 1: Inactivation of lysosomal iron inhibits ferroptosis (Part 1).
From: Activation of lysosomal iron triggers ferroptosis in cancer
a, Chemical synthesis of cLip-1. b, Fluorescence imaging of labelled cLip-1 (1 μM, 2 h) and specific markers of cellular organelles. Scale bars, 10 μm. n = 3 independent experiments. Data are mean ± s.d. c, Fluorescence imaging of labelled cLip-1 (1 μM, 1 h) and LysoTracker. Scale bar, 10 μm. n = 3 independent experiments. Data are mean ± s.d. d-f, Fluorescence imaging of labelled cLip-1 (1 μM, 1 h) and BacMam transduced cells of GFP-labelled cell organelle markers. Scale bar, 10 μm. n = 3 independent experiments. Data are mean ± s.d. g, Fluorescence imaging of labelled cLip-1 in proximal kidney and liver tissues of a Rosa26-CreERT2;Gpx4f/f mouse treated with cLip-1 (0, 10 and 100 mg/ kg/ day; sacrificed 1 h after i.p. injection). Scale bar, 100 μm. h, Bodipy-C11 581/591 flow cytometry of cells treated with Lip-1 (1 µM), cLip-1 (1 µM) and RSL3 (100 nM) for 1 h. i, Viability of cells (top, after 72 h of incubation; middle, incubation for 24 h and cotreatment with RSL3 (500 nM); bottom, incubation for 72 h and cotreatment with 4-OH TAM to knock out Gpx4. Data are mean ± s.d. j, Annexin-V/ Propidium Iodide (A/ PI) staining in cells treated for 24 h. RSL3: n = 2 independent experiments; RSL3 + Lip-1: n = 3 independent experiments; RSL3 + cLip-1: n = 3 independent experiments. k, 1H NMR spectra of Lip-1 and naphthalene titrated with FeCl3. l-n, 1H NMR spectra of Lip-1 titrated with FeCl3 then TFA. o, 1H NMR spectra of Lip-1 titrated with FeCl3 and then sodium deuteroxide (NaOD). k-o, NMR spectra recorded at 310 K in methanol-d4. b, d-f, 1-way ANOVA. AU, arbitrary unit.
