Fig 2: M2-like macrophages express higher levels of Slc7a11 than non-M2-like macrophages.RNA-seq was conducted for four different types of Ly6Clow macrophages; uIRI kidney cortex, sham kidney cortex, uIRI-Can, and Sham-Can. (n = 3 per each kidney sample, n = 4 per each tumor sample). a Principal component analysis (PCA) shows different expression patterns between uIRI-treated samples and Sham-treated samples among Ly6Clow macrophages in the kidney (left) and Ly6Clow macrophages in cancer (right); blue color represents uIRI-treated samples, green color represents Sham-treated samples. b, c Heatmap shows differentially expressed genes. b 441 genes were upregulated and 306 genes were downregulated in Ly6Clow macrophages sorted from the uIRI kidney cortex compared to those from the sham kidney cortex with FDR < 0.001 and log fold change (log FC) more than 2. c 93 genes were upregulated and 59 genes were downregulated in Ly6Clow macrophages sorted from uIRI-Can compared to those from Sham-Can with FDR < 0.05. Representative gene tracks of RNA-seq signals around Ccl22 in kidney samples (d), and Spp1 in tumor samples (e). RNA-seq signals are visualized using Integrative Genomics Viewer (Version 2.11.3) (http://software.broadinstitute.org/software/igv/). f 26 genes were commonly upregulated in Ly6Clow macrophages sorted from the uIRI kidney cortex and uIRI-Can. g Top 20 Gene Ontology (GO) terms enriched among upregulated genes in uIRI kidney included the response to oxidative stress. h Scatter plots show the locus of Slc7a11 among differentially expressed genes in kidney sample (left), and in tumor sample (right). i In qPCR, Slc7a11 expression was upregulated in F4/80lowLy6Clow macrophages compared to F4/80highLy6Clow macrophages both in uIRI kidney cortex (left) and uIRI-Can (right). (n = 3 per each F4/80highLy6Clow macrophage group, n = 4 per each F4/80lowLy6Clow macrophage group; two-sided Mann–Whitney U test). uIRI unilateral ischemia-reperfusion injury, uIRI-Can kidney cancer inoculated into the kidney subcapsule after unilateral IRI, Sham-Can kidney cancer inoculated into the kidney subcapsule after sham operation.

Fig 3: Significant correlation between xCT protein levels in CNS tissues and in vivo PET signal. a Representative Western blot with actin and NaK-ATPase as loading controls (Ctrl). Correlation between xCT protein levels (determined by densitometric Western analysis) and [18F]FSPG uptake (determined by PET quantification) in the b cervical/thoracic spinal cord (SC), c lumbar SC, and d brain. ***p 0.0001–0.001, **p 0.001–0.01, *p 0.01–0.05

Fig 4: NaIO3 (40 mg/kg) induced lipid peroxidation in the RPE monolayer and cell death of photoreceptors after 24 h. (a) mRNA expression of ferroptosis-associated genes in RPE/choroid assessed by real-time PCR (n = 6 per treatment condition). (b) xCT and GPx4 protein expression in RPE/choroid assessed by western blot analysis (n = 3 per treatment condition). (c) Immunofluorescence staining of 4-hydroxynonenal (4-HNE) in RPE monolayer (n = 4 per treatment condition). Triangle: radial cut of flat mount; asterisks: optic nerve. Scale bar = 200 µm in the upper row and 100 µm in the lower row. Abbreviations: veh: vehicle; Zil: Zileuton.

Fig 5: Myrislignan suppressed the progression of GBM in vivo. (a) Representative HE staining of mice brain sections indicated that myrislignan suppressed the growth of GBM. (b) Kaplan-Meier curves of mice survival. (c) The tumor-bearing brains were weighed. Relative weight of tumors in brain was estimated (DMSO/Myris groups—PBS groups). (d, e) Protein levels of p-p65, Slug, and SLC7A11 in mouse tumor were detected by western blot and quantified. The protein samples were extracted from tumor tissues in DMSO and Myris group of intracranial xenograft models. (f) Expression of p-p65, Slug, E-cadherin, and SLC7A11 was evaluated via IHC staining. Scale bar, 50 µm. *P < 0.05; **P < 0.01; ***P < 0.001.