Pretreated samples were analyzed in multiple reaction monitoring mode using triple quadrupole LC-MS/MS (LCMS-8050) and the Cell Culture Profiling Method Package (Shimadzu Corp). This combination enabled us to simultaneously analyze 95 compounds, including basal medium components and secreted metabolites in 17 min. The 95 metabolites and compounds analyzed are listed in table S2. For normalization of the measured area ratio, culture medium was collected every 24 hours, and the cells were refed with fresh medium. The exponential cell growth curve was generated by plotting cell number counted at every medium collection. The amount of secreted metabolite from a single cell in 1 hour was calculated by dividing the area ratio or micromolar measured by LC-MS/MS with the area defined by the cell number curve of the past 24 hours [in area ratio per cell per hour or in nmol (in 2 ml of culture volume) per cell per hour]. LC-MS/MS data for each experiment are summarized in data file S1. The raw LC-MS/MS data are available in data file S2.

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