Lipidomic raw data
Description
Our aim was to compare the lipidome of HK2 cells incubated with IL-6 or oncostatin M (OSM). Our first aim was to determine if AA-phospholipids are enriched in OSM or IL-6 treated cells. There are 3 biological replicate per condition, and 6 conditions: non)-treated 48 h and 72 h; IL-6 48h and 72 h; and OSM 48h and 72h. 100,000 HK-2 cells were spiked with 4.56 μL of internal standard lipid mixture and subjected to lipid extraction at 4 °C . The sample was dissolved in 200 μL of 155 mM ammonium bicarbonate and then extracted with 1 mL of chloroform-methanol (10:1) for 2 hours. The lower organic phase was collected, and the aqueous phase was re-extracted with 1 mL of chloroform-methanol (2:1) for 1 hour. The lower 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 witha TriVersa NanoMate ion source (Advion Biosciences). 5 µL of sample were 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.