Extended Data Fig. 2: The antioxidant effect of lipoproteins is independent of MUFAs and vitamin K.
From: Glycosaminoglycan-driven lipoprotein uptake protects tumours from ferroptosis
a. Proliferation (log2 doublings, 5 days) of indicated GPX4_KO cell lines in vitro either untreated or supplemented with Fer-1 (1 μM), α-toc (10 μM), vitamin K2 (10 μM), or oleic acid (OA, 250 μM). b. Proliferation (log2 doublings, 5 days) of indicated GPX4-deficient cell lines in vitro left untreated or supplemented with α-toc, vitamin D3, or 7-DHC at indicated concentrations. c. Proliferation (log2 doublings, 5 days) of A-498 cells in vitro either treated with ferroptosis inducers ML162 (left) or erastin (right), with or without α-toc, vitamin D3, or 7-DHC at indicated concentrations. d. Cellular lipid oxidation ratio using BODIPY-C11 in A-498 cells untreated or treated with ML162 (125 nM) ± LDL, HDL, α-toc (10 μM), vitamin K2 (10 μM), vitamin D3 (10 μM), CoQ10 (10 μM), OA (250 μM) or Fer-1 (1 μM). e. Cellular lipid oxidation ratio using BODIPY-C11 in A-498 cells ± LDL, HDL, α-toc (10 μM), vitamin K2 (10 μM), OA (250 μM) or Fer-1 (1 μM). f. Cellular lipid oxidation ratio using BODIPY-C11 in Karpas299 cells untreated or treated with ML162 (125 nM) ± LDL, HDL, α-toc (10 μM), vitamin K2 (10 μM), vitamin D3 (10 μM), vitamin A (10 μM), CoQ10 (10 μM), OA (250 μM) or Fer-1 (1 μM). g. Schematic of genes required for cellular utilization of antioxidant lipids carried by lipoproteins. h. Immunoblot of ACSL3 and AIFM2 in the indicated cell lines transduced with a sgControl, sgACSL3 or sgAIFM2. GAPDH is the loading control. i, j. Cellular lipid oxidation ratio using BODIPY-C11 in Karpas299 (left) or A-498 (right) cell lines transduced with sgACSL3 (i) or sgAIFM2 (j) versus sgControl under GPX4 inhibition (ML162: 75 nM for Karpas299, 175 nM for A-498) and HDL supplementation. For all LDL and HDL experiments, 50 ug/mL was used. a-c, Bars represent mean ± s.d.; d-f, i, j, bars represent median; a-f, i, j, n = 3 biological replicates. Statistics by two-sided unpaired t-tests as indicated or compared to untreated cells (a-c, e) or ML162 treated cells (d, f). For gel source data, see Supplementary Fig. 1.
