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TMY85 cells were cultured overnight in YPD medium containing 5 μM PHS. The cells were washed and then diluted (0.1 D600 unit/ml) in YPD medium containing 5 μM PHS (B), 5 μM DHS (C) or 5 μM SPH (D), and incubated for 18 h at 30 °C. Lipids were extracted, hydrolysed by methanol/HCl, derivatized with OPA and analysed by reversed-phase HPLC. Details are provided in the Experimental section. (A) PHS, DHS and SPH standards.
