Extended Data Fig. 1: Dose-response of D-PUFAs, peroxisome and mitochondrial staining of D-PUFA-treated cells, and mitochondrial/ER quantification of D-PUFAs.
From: Identification of essential sites of lipid peroxidation in ferroptosis
a. Dose-response curves of HT-1080 cells pre-treated with varying concentrations of D-PUFAs and then treated with FINs. Data are plotted as mean ± SEM, n=3 biologically independent samples. b. HT-1080 cells treated for 24 hours with 20 µM ARA-d6 and CellLight Peroxisome-GFP, and imaged by fluorescence and SRS imaging. c. HT-1080 cells treated for 24 hours with 20 µM DHA-d10, and then stained with MitoTracker Red CMXRos and imaged by fluorescence and SRS imaging. d. High resolution SRS imaging of HT-1080 cells treated for 24 hours with 20 µM DHA-d10 with DGAT inhibitors PF-06424439 (1 µM) and A922500 (1 µM), then stained with MitoTracker Red CMXRos and ERTracker Green and imaged by fluorescence and SRS imaging. e. Quantification of arachidonic acid-d11 in mitochondrial and ER fractions isolated from HT-1080 cells treated at 20 µM for 24 hours. Values determined by high resolution mass spectrometry and plotted as normalized to internal standard. Data are plotted as mean of three biological replicates ± SEM. f. Western blotting of mitochondrial and ER fractions stained for PDI as ER marker and Cytochrome C as mitochondrial marker, representative of four experiments. Results indicate no mitochondrial contamination of ER fraction, but indicate ER contamination of mitochondrial fraction.
