Fig. 1
Exosome-packaged RNA from CAFs enhances the glutamine uptake and progression of LUAD cells. A, B Representative transcriptome analysis of the Reactome pathway (A) and GSEA (B) in the A549 cells co-cultured with CAFs compared with those with NFs (n = 3). B Reactome pathway-based GSEA revealed an enrichment of “Metabolism of amino acids and derivatives” in the A549 cells co-cultured with CAFs compared to NFs co-cultured with A549 cells (n = 3). C OCR of A549 cells with indicated treatments (n = 3). D ATP production of A549 cells with indicated treatments (n = 3). E Schematic and heatmap of metabolomic experiments for conditioned media of indicated cells. F–L A549 cells were treated with exosomes or CM of CAFs and then used for the indicated experiments. anti-CD81 antibody was used for depleting exosomes in the CM of CAFs. F OCR of A549 cells with the indicated treatments (n = 3). G ATP production of A549 cells with indicated treatments (n = 3). H Extracellular glutamine levels of A549 cells cultured for 4, 8, and 12 h in 25 mM glucose DMEM (containing 1 mM pyruvate and 4 mM glutamine) with the indicated treatments (n = 3). I 3H-glutamine uptake (n = 3). J Glutamine-derived TCA cycle intermediates in A549 cells with indicated treatments (n = 3). K, L RTCA proliferation, migration (lower), and invasion (upper) assays and ATP production of A549 cells with indicated treatments. M Extracellular levels of glutamine of A549 cells with the indicated treatments (n = 3). N RTCA proliferation assays, migration (lower), and invasion (upper) assays of A549 cells with indicated treatments (n = 3). For C, D and F–N, Means ± s.d. are shown, and independent sample t-tests determined P values. * P < 0.05, ** P < 0.01, *** P < 0.001. UT, cancer cells without any treatment; CM, conditioned medium; Exos, exosomes; GSEA, gene set enrichment analysis; RTCA, Real-time xCELLigence analysis; LUAD, lung adenocarcinoma