Lipidomic raw data

Male C57 mice were administered 0.5 mg/kg of dACSL4@BAmp-TK12 (dosage calculated based on dACSL4) intranasally for three consecutive days. One day after the final administration, the brains were collected, minced, and homogenized. For lipid extraction, a mixture of methanol: methyl tert-butyl ether: water (3: 10: 2.5, v/v) was added, and the sample was vortexed three times. The mixture was centrifuged at 3,000 rpm for 15 min at 4°C. The upper organic phase was transferred to another new glass tube, dried under nitrogen, and resuspended before the test. The lipids levels were detected by a Q Exactive LC-MS/MS (Thermo Fisher Scientific) system and quantitatively analyzed using the area external standard method according to the ratio of the relative amounts of response area of lipids. For comparison between different groups of cells, lipid expression was homogenized by the values of all lipid expression levels so that they conformed to the standard normal distribution. During the lipid extraction process, it was corrected according to the total protein concentration in the corresponding sample to ensure that the number of cells in the different samples was the same. Hence, the expression of lipids would not be affected by the sample size.
There are 3 biological replicates per condition. Cells were incubated with 1 μM dACSL4 or an equivalent volume of DMSO for 12 h, then harvested by trypsinization (500 μL trypsin), diluted with 500 μL PBS, transferred to 1.5 mL tubes, and centrifuged at 800 rpm for 5 min at 4°C. The organic phase was collected and evaporated in a SpeedVac vacuum concentrator. Lipid extracts were dissolved in 100 μL of infusion mixture consisting of 7.5 mM ammonium acetate dissolved in propanol:chloroform:methanol [4:1:2 (vol/vol)].
Samples were analyzed by direct infusion in a QExactive mass spectrometer (Thermo Fisher Scientific) equipped with a TriVersa NanoMate ion source (Advion Biosciences). 2 µL of sample was infused with gas pressure and voltage set to 1.25 psi and 0.95 kV, respectively.
All data was acquired in centroid mode. All lipidomics data were analyzed with the lipid identification software, LipidXplorer 29. Tolerance for MS and identification was set to 2 ppm. Data post-processing and normalization to internal standards were done manually in Excel.