Extended Data Fig. 3: GC–MS detection of OAA 3-TBDMS and analysis of glucose uptake.

a. (left) Scan of m/z 417 of pure ¹²C OAA standard derivatized with DMF-MTBSTFA. (middle) Mass spectrum of pure ¹²C OAA standard, derivatized with DMF-MTBSTFA. m/z 417 corresponds to OAA 3-TBDMS M-57 fragment. (right) Standard curve showing a positive, linear relationship between OAA amount and m/z 417 peak intensity of pure OAA standard. b. (left) Scan of m/z 417 of H1299 cellular metabolites labelled with ¹³C glucose, extracted with 1:1 methanol:water containing 5 mM ninhydrin, with or without ¹²C OAA spike. (middle) Isotopomer distribution of OAA (m/z 417) from H1299 cellular metabolites labeled with ¹³C glucose, extracted with 1:1 methanol:water containing 5 mM ninhydrin, with or without ¹²C OAA spike. (right) Ion abundance, normalized using IsoPat2, of isotopomer distribution data from (middle). c. Relative glucose uptake of H1299 (left) and RPMI 8226 (right) cells treated with vehicle or 1 mM diethyl-ester OAA. d. Relative glucose uptake of H1299 (left) and RPMI 8226 (right) cells treated with vehicle or 10 mM aspartate. Data is represented as the mean and error bars represent the standard deviation from n = 4 for (c), (d) of biologically independent replicates. Results in (a) are representative experiments of three independent replicates in vitro. Results in (b) are representative experiments of 3 biologically independent replicates. P values were determined by a two-tailed Student’s t-test.