Fig. 5

Metabolite changes in glycolysis and the TCA cycle associated with methionine stress. a MB468 and MB468res-R8 cells were cultured in Met+ or Met-Hcy+ media over the course of 24 h and analyzed by untargeted metabolomics as described for Fig. 4. Heatmaps represent fold changes of metabolites in glycolysis and the tricarboxylic acid (TCA) cycle as compared to the Met+, time-zero sample. Metabolites are color filled by fold change: yellow indicates a decrease in metabolite abundance and blue indicates an increase as compared to time-zero control. b Schematic of glycolysis and the TCA cycle. Metabolites with gray arrows are pointing to line graphs of the corresponding metabolite abundances in MB468 (blue line with squares) and MB468res-R8 (black line with circles) cells. Peak height reflecting relative metabolite abundance is indicated on the y-axis and hours in homocysteine are indicated on the x-axis. Error bars represent standard deviation. Metabolites included in the heatmaps (a) are outlined in a dashed box. c MB468 and MB468res-R8 cells cultured in Met+ media for 12 h or Met-Hcy+ media for 2, 4, 8, and 12 h were analyzed by the Seahorse Bioscience XF analyzer. MB468 (blue line with squares) and MB468res-R8 (black line with circles) basal extracellular acidification rates (ECAR) are shown normalized to the Met+-treated sample. Error bars represent normalized standard deviation, n = 4 replicates; statistical differences between Met+ media and Met-Hcy+ media treatments are indicated by asterisk where p ≤ 0.002