Fig. 3

Aspirin regulates proteins involved in central carbon metabolism. a ATF4 target genes highlighted in proteomic data by IPA analysis. Fold change of proteins in both long-term (52 weeks) 2-mM and 4-mM aspirin conditions, relative to control (n = 3 biological replicates). IPA analysis shows a predicted overall inhibition of ATF4 signalling with long-term 4mM aspirin (p = 0.0116, z score =  − 2.894). b Overview of key enzymes involved in glutaminolysis and their average fold changes with long-term 4-mM aspirin treatment in the proteomic data. BCAAs, branched-chain amino acids; OAA, oxaloacetate, PHP, phosphohydroxypyruvate. Created with BioRender.com. c Metabolite abundance relative to cell number of alanine and aspartate in long-term 4-mM aspirin-treated cells compared to control. Error bars represent SD (n = 6 technical replicates). Asterisks refer to p values obtained using t tests ***p < 0.001, ****p < 0.0001. Created with BioRender.com. d Immunoblotting for a selection of metabolic enzymes highlighted in proteomic data, with long-term aspirin treatment (ns, non-specific). Representative of at least 3 independent experiments. α-tubulin is used as a loading control. e Average fold changes in the proteomic data with long-term 4-mM aspirin compared to control, including central carbon metabolism genes that showed significant regulation (p < 0.05, fold change > 1.4) in both long-term 2mM and 4mM aspirin. Created with BioRender.com