Strains were cultured at 30°C unless indicated otherwise. Optical density at 600 nm (OD600) was measured with a Hitachi U‐2000 double‐beam spectrophotometer. The WT strain was pre‐cultured in SD without choline (C−), whereas cho2opi3 and derived triple mutants were pre‐cultured in SD with choline (C+) to the mid‐log phase of growth (OD600 ˜0.45–1.2). Cells transformed with plasmids were grown in selective SD drop‐out media. To deplete phosphatidylcholine, cho2opi3 cells were collected by centrifugation or filtration where indicated, washed thoroughly with pre‐warmed SD C− (30°C), and transferred to fresh SD C− at the initial OD600 indicated (Boumann et al, 2006 (link)). SH80 and SH85 were pre‐cultured in YPD. Doubling time (DT) was determined based on OD600 values at different time points according to:
DT = ln2/μmax with μmax the growth rate during exponential growth, μmax = (lnOD600t2 ‐ lnOD600t1)/(t2‐t1).
DT = ln2/μmax with μmax the growth rate during exponential growth, μmax = (lnOD600t2 ‐ lnOD600t1)/(t2‐t1).