Extended Data Fig. 7: Deuterium tracing to interrogate paralog pathway-mediated metabolic compensations.
(a) Stable-isotope tracing map elucidating the transfer of [2H] and enrichment of downstream metabolites from stable-isotope substrates 3-[2H]-glucose (blue), to trace [2H]-NADPH and cytosolic MTHFD1 activity, and 4-[2H]-glucose (orange) to trace [2H]-NADH and mitochondrial MTHFD2 activity. All four ovarian cancer cell-lines were cultured separately in each of the two tracers, and intracellular metabolites were analyzed after 3, 6, and 24 hours of culture with labeled media. Data are presented as mean ± s.d. (n=3). (b) Stable-isotope tracing map elucidating the transfer of 2H and enrichment of downstream metabolites from stable-isotope substrate 2,3,3-[2H]-serine (green) to dissect bidirectionality of SHMT2 and MTHFD2 fluxes in mitochondria and SHMT1 and MTHFD1 in cytosol. Intracellular metabolites were analyzed after 3, 6, and 24 hours of culture with labeled media. Data are presented as mean ± s.d. (n=3). Data analyzed using ordinary 2-way ANOVA with Dunnett’s correction for multiple comparisons.
