Extended Data Fig. 1. TRCs resist radiotherapy and chemotherapy via evading ferroptosis.
a-b, HONE1 cells and HK1 cells were seeded in 3D fibrin gels for 5 days of culture. The colony size of the tumor spheroid was presented (a). The colony sizes at different time points were quantified and the colony size on d1 was set to 1 (b). Bar, 50 mm. n = 6. c, Immunoblots showing the expression of cancer stem cell markers (pSTAT3 and SOX2) in TRCs and bulk cells. d, RNA-seq was performed on HONE1 TRCs and bulk cells. The gene set enrichment analysis (GSEA) shows that multiple stem cell-like transition pathways were enriched in the TRCs group. Two-sided P values were calculated with Kolmogorov-Smirnov test and multiple-comparison adjusted q values were calculated using Benjamini-Hochberg (BH) method. e, The HONE1 TRCs and bulk cells (5 cells or 100 cells) were injected into the mammary fat pads of NSG mice. Eight weeks later, the tumor formation was recorded. n = 12. f-g, HONE1 TRCs and bulk cells were treated with cisplatin (Cis) with or without Fer-1. The content of ferroptotic cell death signal, PE (38:4)-OOH (f) and PE (40:4)-OOH (g) was measured. h-j, Dose-dependent toxicity of RSL3 (h), FIN56 (i) or FINO2 (j) induced cell death of HK1 cells. k, Principal component analysis (PCA) of oxygenated phospholipids in HONE1 TRCs and bulk cells. Each point represents a sample. l, Heatmap showing the relative changes in RSL3-induced oxygenated PE and PC molecular species in HONE1 TRCs and bulk cells. Row z-scores were obtained from averaged values of the content of lipid species. Data are shown as mean ± SD (b, f-g), one-way ANOVA (f-g). n = 3 independent experiments. One of three experiments is shown (c). ns, not significant.