Fig. 4
Metabolomic analysis of H460 and MCF7 cell lines grown in 2D and 3D culture conditions. A Heatmap of 66 significantly altered metabolites in H460 and MCF7 cell lines in 2D vs 3D conditions. B Dot plots showing the 7 metabolites with the same trend of variation in all 3D vs 2D cultures of both cell lines. C KEGG pathway enrichment analysis of glycolysis/gluconeogenesis, showing in red ALDOC and ENO2 upregulation at both gene and protein levels. D-E Intracellular glucose and L-lactic acid amounts measured by luminometric assays and reported as relative light units (R.L.U.); quantification of L-lactic acid within the culture media (extracellular) performed by emogas analysis and expressed as mmol/l in H460 2D, H460 3D_SM, H460 3D_ FBSlow, MCF7 2D, MCF7 3D_SM, and MCF7 3D_ FBSlow. All the experiments were carried out in triplicate and results are presented as mean ± SD. p-value: * < 0.05, ** < 0.01, *** < 0.001. ns: not significant