Fig. 7. Lipid peroxidation by αESA is promoted by ACSL1.

a Western blot showing ACSL1 protein expression in control or ACSL1-silenced cells. The efficiency of the siRNA knockdown was independently confirmed using quantitative PCR (qPCR) (Supplementary Fig. 4b). b Bar charts showing the overall amount of 18:3-containing TAGs in BT-549 cells under the specified condition (n = 3 biological replicates for this and subsequent bar charts). Cells were treated for 8 h with 25 μM αESA or vehicle. Treatments were added 72 h after transfection with a pool of either ACSL1-targeting or non-targeting siRNA. In this and subsequent panels, mean values ± standard deviation are presented, and p-values are shown above comparator bars (two-sided Student’s t-test). c Volcano plot showing log₂(fold change) and significance (log₁₀(1/p)) for oxidized phospholipids in BT-549 cells incubated for 8 h with 25 μM αESA compared to cells incubated with vehicle (methanol). Each lipid species is colored according to phospholipid class. BMP = bis(monoacylglycero)phosphate, PS = phosphatidylserine, PE = phosphatidylethanolamine, PG = phosphatidylglycerol, PI = phosphatidylinositol, PC = phosphatidylcholine, CL = cardiolipin. The arrow indicates a lipid species for which a fold change could not be computed because it was only detected after αESA treatment. d Bar charts showing the amounts of di-oxygenated PE species that were significantly increased by αESA treatment compared to vehicle-treated controls. e Volcano plot depicts log₂(fold change) versus log₁₀(1/p-value) for oxidized phospholipids in BT-549 cells transfected with a pool of ACSL1 siRNA and incubated with 25 μM αESA for 8 h compared to αESA-treated cells transfected with non-targeting siRNA. The set is limited to the 51 oxidized phospholipid species that were significantly changed by αESA treatment as shown in c. Bar charts showing the amount of a (f), di-oxygenated and (g), mono-oxygenated TAG species that were significantly increased by αESA treatment in an ACSL1-dependent manner. Source data are provided as a Source Data file.