Extended Data Fig. 4: PA correlates with areas of high MYC and anticorrelates with lactagenic metabolism.

a-c, Labeled compounds utilized in DEFFI or NanoSIMS imaging as indicated. d, Aligned post DEFFI fluorescent microscopy and glutamate M + 5 isotopologues measured by DEFFI after [¹³C₅]glutamine infusion in two repeat runs (Run 1 is displayed in Fig. 3a). e,f, Post DEFFI fluorescent microscopy of WM^Mix tumors, shows increased PA in WM^High regions. The bottom panels are binarised representations of the labeled proxy compounds (lactate M + 3 isotopologue for lactagenic metabolism, malate M + 1 isotopologue for increased Krebs cycle, glutamate M + 5 for glutamine uptake) showing Krebs cycle activity corresponds with higher PA and WM^High areas. Ion intensities were corrected for natural abundance from individual metabolites and for each pixel the metabolite with the higher value after maximal intensity normalization was depicted. Note that for illustrative purposes some panels are also displayed in Fig. 3c,d. g,h, Table showing the pairwise correlation between different detected labeled compounds and PA. The top value represents the correlation and the bottom value the p value (n = 3 tumors from independent animals). i, j, Cell-wise quantification of ¹³C/¹²C and ¹⁵N/¹⁴N ratios in WM^Mix tumors (run 2 and run 3 each representing a tumor from an independent biological replicate, run 2: ¹³C/¹²C, p = 3.53e⁻⁵; ¹⁵N/¹⁴N, P = 0.141; run 3: ¹³C/¹²C, P = 3e⁻⁸; ¹⁵N/¹⁴N, P = 1.14e⁻¹⁰) k,l, Cell-wise quantification of ¹³C/¹²C and ¹⁵N/¹⁴N ratios in WM^Mix tumors stratified by BrDu incorporation status (n = 3 tumors from independent animals, ((k) run 1: Brdu⁻, P = 1.2e⁻¹¹; BRDU⁺, P = 0.0312; run 2, P = 0.23; run 3, P = 5.94e⁻⁹; (l): run 1, P = 0.0001; run 2, P = 0.0004; run 3, P = 5.33e⁻⁷). All box and whisker plots represent the following: Line: median, box: IQR, whiskers: 1.5x IQR limited by largest/smallest NEV. Significance was calculated with an unpaired two-tailed t-test. P value: ****<0.00001. See also Fig. 3.