Fig. 2

GlaB treatment significantly altered (*) the metabolic profile of GL261 cells. a Metabolomic phenotyping of cell lysates (left panel, endo-metabolome) and respective growth media (right panel, exo-metabolome) after 12 h, 24 h and 48 h of GlaB treatment. Score plots of PCA: PC1 vs PC2. The 6-group discrimination accuracy values are also reported. In the score plots, each dot represents a different sample, and each color represents a different group of samples: blue dots, 12-CTR; red dots, 24-CTR; purple dots, 48-CTR; green dots, 12-GLAB; orange dots, 24-GLAB; cyan dots, 48-GLAB. b Schematic representation of glycolysis and TCA cycle in GL261 cells. c Box plots showing the altered intracellular glycolytic metabolites over time. d Box plots showing the altered extracellular glycolytic metabolites over time. e Box plots showing the altered intracellular TCA-metabolites over time. f Box plots showing the altered intracellular TCA-related metabolites over time. g Box plots showing the altered extracellular TCA-metabolites over time. Data are expressed as mean ± se, n = 5 *p < 0.05 vs respective control. h Western blot analysis of GlaB treated (5 μm) GL261 cells at indicated time point for AMPK phosphorylation. Quantification of protein expression by densitometry analysis of the bands. Data are expressed as fold increase over control of phospho/total protein ratio of Ampk. Statistical analysis: One-way ANOVA followed by Dunn’s post hoc test *p ≤ 0.05. On the right, representative western blots