Fig. 3
HIF-2α selectively enriches polyunsaturated lipids. a Heatmap representing the relative lipid abundances in indicated cell lines. The abundance of each lipid species is normalized to the mean of that in the EPAS1+/+ 786-O WT cells and the ratios are log2 transformed. The lipids are grouped by classes, and within each class, the lipid species are ordered first with increasing carbon number, then with increasing unsaturation levels. Abbreviations: CE, cholesterol ester; Cer, ceramide; MAG, monoacylglycerol; DAG, diacylglycerol; TAG, triacylglycerol; LPC, lysophosphatidylcholine; LPE, lysophosphatidylethanolamine; PC, phosphatidylcholine; PE, phosphatidylethanolamine; ePC, (vinyl ether-linked) PC-plasmalogen; ePE, (vinyl ether-linked) PE-plasmalogen; PI, phosphatidylinositol; PS, phosphatidylserine; SM, sphingomyelin. Blue: down-regulated relative to the WT cells, red: upregulated relative to the WT cells. The wave-like pattern in the TAG class corresponds to the more significant losses in the polyunsaturated fatty acyl (PUFA)-TAGs than saturated/monounsaturated fatty acyl (SFA/MUFA)-TAGs in response to HIF-2α-depletion. S, ferroptosis-sensitive; (R), ferroptosis-resistant. b Volcano plots showing the changes in TAGs grouped as PUFA-TAGs (red fill) and SFA/MUFA-TAGs (white fill) between the indicated cell lines. n = 3, two-tailed t-test. c Volcano plots showing changes in PE and ePE lipids grouped as PUFA-PE/ePEs (red fill) and SFA/MUFA-PE/ePEs (white fill) between the indicated cell lines. d Bar graph representing the relative abundances of the indicated PUFA-PE/ePE lipids in the labeled groups. Log2 fold changes relative to 786-O WT cells are presented for each condition. n = 3, error bars: ±s.d. e Bar graph representing the relative abundances of the indicated free fatty acids grouped as PUFAs or SFA/MUFAs in the indicated conditions. nd, not-detectable under the experimental condition. n = 3, error bars: ±s.d. f Viability curves of 786-O-Cas9 and 769-P-Cas9 cells expressing control (sgNC) or EPAS1-targeting sgRNAs, first treated with BSA or PUFA (arachidonic acid, C20:4) for 3 days, then treated with indicated concentrations of ML210 or RSL3 for 48 h. Representative plot of experiments repeated twice. g The ratios between PUFA-PE/ePE and total PE/ePE (left), and between PUFA-PC/ePC and total PC/ePCs in ccRCC tumor samples (n = 49; red) and the matched normal tissues (n = 49; gray) from previously reported lipidomics datasets. Student’s T-test, ***p < 0.001
