Fig. 7: Oleic and palmitic acid supplementation in HepG2 increases PG-PUFA2 leading to ferroptosis susceptibility. | Cell Death & Differentiation

Fig. 7: Oleic and palmitic acid supplementation in HepG2 increases PG-PUFA2 leading to ferroptosis susceptibility.

From: Ferroptosis is a targetable detrimental factor in metabolic dysfunction-associated steatotic liver disease

Fig. 7

To study the effect of lipid accumulation on ferroptosis sensitivity, HepG2 cells were exposed to glucose, insulin, three cytokines and different species of fatty acids, i.e., oleic and palmitic acid (OA/PA), arachidonic acid (AA 20:4, ω-6 PUFA) or docosahexaenoic acid (DHA 22:6, ω-3 PUFA). A Experimental design of fatty acid incubation and ferroptosis induction with GPX4 inhibitor ML162. B Neutral lipids measured with AdipoRed. C Percentage of cell death (with SytoxGreen) at increasing ML162 concentrations. After fitting non-linear regressions per condition, best-fit half maximal effective concentration (EC50) values were compared using Aikake Information Coefficient and one-way ANOVA with post-hoc testing. D Representative images of C11-BODIPY (581/591) dye in HepG2. Oxidized dye (green) indicates lipid radical oxygen species 2 h after ML162, as opposed to the normal reduced form (red). E Gene set enrichment analysis (GSEA) plots for gene sets “ferroptosis defenses” and “GSH” in HepG2 treated with OA/PA compared to solvent control (Control), with normalized enrichment score (NES) (n = 6). F Percentage of phosphatidylglycerol (PG) esterified with one or two polyunsaturated fatty acids, i.e., PG-PUFA and PG-PUFA2. Percentage of phosphatidylcholine and phosphatidylethanolamine with two PUFA, i.e., PC-PUFA2 and PE-PUFA2, respectively (n = 3). Data pooled from three independent experiments, performed in triplicate, except for C11-BODIPY (581/591). *p < 0.05; **p < 0.01; ***p < 0.001. One-way ANOVA with post-hoc test.

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