Profinder b 0

Extracted metabolites were separated on a Cogent Diamond Hydride Type C column (gradient 3) (Microsolve Technologies) and the mobile phase consisted of solution A (ddH2O with 0.2% formic acid) and solution B (acetonitrile with 0.2% formic acid). The mass spectrometer used was an Agilent Mass 6230 time of flight (TOF) coupled with an Agilent 1290 liquid chromatography (LC) system. Detected ions were deemed metabolites on the basis of unique accurate mass-retention time identifiers for masses exhibiting the expected distribution of accompanying isotopologs. The abundance of extracted metabolites was measured using Agilent Qualitative Analysis B. 08.00 software and Profinder B.0.800 software (Agilent Technologies) with a mass tolerance of <0.005 Da. All data obtained by metabolomics were the average of three independent samples of each condition tested. Analysis was performed on metabolite intracellular concentration per milligram of total proteins (IC/mg) taken from GC-MS. The normalized metabolomic data used in this study are listed in Table S2. The variance of metabolome data was analyzed by 2D Principal Component Analysis and heatmap using Clustvis (https://biit.cs.ut.ee/clustvis/) (Metsalu and Vilo, 2015 (link)). Statistical significance was calculated using GraphPad PRISM using a two-way ANOVA with HSD Tukey post-hoc analysis.