Extended Data Fig. 2: Peroxisomes contribute to ferroptosis sensitivity in renal and ovarian carcinoma cells.
From: Plasticity of ether lipids promotes ferroptosis susceptibility and evasion
a, Fluorescent imaging analysis of peroxisome abundances, reported by PTS1–GFP signal, in OVCAR-8 (top) and 786-O (bottom) cells expressing sgNC, or PEX3, PEX10-targeting sgRNAs. Scale bar, 50 μm. Peroxisome quantifications are shown on the right as violin plots. OVCAR-8 sgNC, n = 295; PEX3 sg1, n = 318; PEX10 sg1, n = 301; 786-O sgNC, n = 604; PEX3 sg1, n = 617; PEX10 sg1, n = 543. Lines in violin plots indicate median and quartiles. b, Fluorescent imaging analysis of peroxisome abundances, reported by PTS1–GFP signal, in OVCAR-8 and 786-O cells expressing sgNC, or PEX12-targeting sgRNAs. Scale bar, 50 μm. Peroxisome quantifications are shown on the right as violin plots. OVCAR-8 sgNC, n = 1,032; PEX12 sg1, n = 259; PEX12 sg2, n = 444; 786-O sgNC, n = 1,139, PEX12 sg1, n = 623; PEX12 sg2, n = 1,326. Lines in violin plots indicate median and quartiles. c, Immunoblot analysis showing the PEX10 levels in OVCAR-8 cells expressing sgNC or PEX10-targeting sgRNAs. COX-IV was used as a loading control. Representative result of experiment performed in duplicate. See Supplementary Information for uncropped immunoblot images. d, Viability curves of 786-O cells expressing negative control (sgNC) or sgRNAs targeting PEX3, PEX10 or PEX12 treated with the indicated concentrations of ML210 or RSL3 for 48 h. n = 4 biologically independent samples. Representative result of experiment performed in triplicate. e, Viability curves of PEX3 sg1-expressing OVCAR-8 cells rescued with sgRNA-resistant mouse Pex3 cDNA respectively and being treated with indicated concentrations of ML210 or RSL3. In this experiment, cellular viability was measured at 24 h of treatment, at which time point the PEX3-sg1-only cells were not yet dying. n = 4 biologically independent samples. Data from an experiment performed once. f, Viability curves of PEX10 sg1-expressing OVCAR-8 cells rescued with sgRNA-resistant mouse Pex10 cDNA respectively and being treated with indicated concentrations of ML210 or RSL3. In this experiment, cellular viability was measured at 24 h of treatment, at which time point the PEX10 sg1-only cells were not yet dying. n = 4 biologically independent samples. Data of experiment performed once. For viability curves, data are mean ± s.d.
