Fig. 1
Clear-cell carcinoma cells are intrinsically sensitive to GPX4 inhibition-induced ferroptosis. a Volcano-plot showing compound sensitivity comparison by normalized area-under-curve (AUC) values between clear-cell carcinoma (CCC) cells (n = 26) and other solid tumor cancer cell lines (sCCL) (n = 634) in CTRP. Cpds, compounds. b Chemical structures of GPX4 inhibitors ML210, RSL3, and ML162. c Scatterplot of AUCs for ML210 in sCCL (blue), CCC (red) or cancer cell lines from each tissue. Tissue types are ordered by the average AUC values. Abbreviations: CNS, central nervous system; UAT, upper aerodigestive tract; a_ganglia, autonomic ganglia. Mann–Whitney–Wilcoxon test, ****p < 0.0001. d Viability curves for the indicated cells treated with ML210 or RSL. n = 4. Representative plot of experiments repeated three times. e Viability curves for the indicated cells treated with ML210 or RSL3 plus indicated DMSO, liproxstatin-1 (Lip-1) or ferrostatin-1 (Fer-1). Representative plot of experiments repeated three times. f Fluorescent images of BODIPY-C11 stained 786-O, 769-P, and BFTC909 cells treated with ML210 plus DMSO or Lip-1 for the indicated time periods. Scale bars: 10 µm. g Immunoblot showing GPX4 protein levels in GPX4+/+ 786-O or GPX4−/− clones 3A2 and 3A7. h Viability curves for WT 786-O, 3A2, and 3A7 over a 2.5-day time course after Fer-1 removal. Representative plot of experiments repeated three times. i Tumor volume measurements of subcutaneous xenografts of WT 786-O or GPX4−/− 3A2 cells (n = 10 mice, two tumors per mouse). GPX4−/− tumor-bearing mice were divided to a Lip-1 treated group and a vehicle-treated group. Treatment lasted for the first 10 days. Two-tailed t-test, ***p < 0.001, ****p < 0.0001. j Immunoblot showing HIF-2α protein levels in primary human ccRCC cell lines CCLF_KIPA_0001_T, CCLF_KIPA_0002_T, and normal renal-cell culture CCLF_KIPA_0001_N matched with CCLF_KIPA_0001_T. k Viability curves for indicated cell lines treated with ML210 or RSL3 plus DMSO or 0.5 µM of Lip-1. n = 4. Representative plot of experiments repeated twice. l Viability curves for ovarian clear-cell carcinoma (OCCC) cell lines ES-2, OVISE, and TOV21G, and high-grade serous carcinoma (HGSC) line OV-90 treated with ML210 (left) or RSL3 (right). n = 4. Representative plot of experiments repeated twice. Error bars: ±s.d. β-Actin was used as loading controls for immunoblots
