Fig. 3: ATM deficiency disrupts glycolytic capacity and reprograms to glutamine dependence in the cerebellum.

a Metabolite set enrichment analysis of commonly downregulated metabolites enriched in cerebellar tissues of 3- and 6-mo Atm-KO mice (n = 6, Globaltest with Bonferroni correction¹⁷³). b Integrated gene-metabolite network analysis in cerebellar tissues harvested from 6-mo Atm-WT and KO mice. Significantly changed KEGG metabolic genes and metabolites were mapped to Reactome pathways (n = 4 for transcriptome; n = 6 for metabolome, Globaltest with Bonferroni correction¹⁷³). c Pie chart illustrating the changes in proportions of glucose, glutamine and fatty-acid dependence in acute ex vivo cerebellar culture (n = 3). Original data is shown in Supplementary Fig. 5g. d Left: schematics of ¹³C₆-U-glucose carbon flow. Right: mass isotopologue analysis of lactate, glycerol-3-phosphate (G3P), α-ketoglutarate (α-KG) in ex vivo cerebellar cultures [n = 16, one-way ANOVA for all except for (1) “M + 0 Lactate” where Kruskal-Wallis test was used]. e Left: schematics of ¹³C₅-U-glutamine carbon flow. Right: mass isotopologue analysis of γ-Aminobutyric acid (GABA), oxaloacetate (OAA), palmitate in ex vivo cerebellar cultures [n = 16, one-way ANOVA for all except for (1) “M + 0 GABA”, (2) “M + 4 OAA”, (4) “M + 0 GABA” and (6) “M + 0 and M + 2 Palmitate” where Kruskal-Wallis test was used]. f Representative immunoblots on importin α3-PKM2 interaction and PKM2 protein quaternary structure status (n = 16). Quantifications presented in Supplementary Fig. 5i. g Representative immunoblot images on subcellular localization of PKM2 (n = 16). Quantification presented in Supplementary Fig. 5j. h Schematics of possible fates of PKM2 cytosolic dimer/monomers. i Representative blot images of RNA-immunoprecipitation assay for the interaction between PKM2 and the IRES region of the c-Myc mRNA harvested from human patient fibroblasts (n = 10). Quantification presented in Supplementary Fig. 6b. j Representative immunoblots showing levels of c-Myc protein in human patient fibroblasts (n = 12). Quantification presented in Supplementary Fig. 6c. k Schematics of human c-Myc consensus binding site sequences. Representative ChIP-qPCR blots showing levels of c-Myc binding to glutaminase-2 (GLS2) promoter in human patient fibroblasts (n = 10). Quantification presented in Supplementary Fig. 7d. Unless otherwise specified, all ex vivo glargine and TEPP-46 treatments were performed in 100 nM for 2 h. Values represent the mean ± s.d. N presents biological replicates. Source data are provided as a Source Data file.