Nature research paper: A non-canonical vitamin K cycle is a potent ferroptosis suppressor
HepG2 cells were seeded on 6 well plates. On the next day, medium was replaced with fresh medium with or without warfarin . On the following day, cells were incubated in the presence or absence of MK4 for 7 h. After washing with PBS three times, cells were trypsinized and collected. Cell pellets were suspended in 400 µl PBS, supplemented with 20 µl of MK4-d as internal standard, and sonicated for 30 s with a sonication probe .
. Four-hundred microlitres ethanol and 1.2 ml hexane were added to the cell lysate followed by shaking for 5 min. Samples were centrifuged at 1,000for 5 min, and the upper organic layer was collected. Re-extraction of the remaining aqueous phase was performed by addition of 150 µl ethanol and 450 µl hexane with subsequent vortexing. Samples were centrifuged at 1,000for 5 min.
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