Fig 2: Increase of Antimicrobial LC3-Associated Immune Defense in Butyrate Macrophages(A) Extracellular acidification rate (ECAR) measured at steady state in control and butyrate macrophages. Data represent the mean of nine biological replicates from three independent experiments.(B–D) Quantification of glycolysis (B), glycolytic capacity (C) and glycolytic reserve (D).(E) Heatmap of metabolites that were significantly higher or lower in control and butyrate macrophages as detected by mass-spectrometry (left panel). Results are from five healthy donors. Right panel: Fold change of all significantly higher or lower metabolites in control macrophages (black closed circles) and butyrate macrophages (red closed circles).(F) AMPK phosphorylation (Thr172) measured by ELISA (n = 4 individual donors).(G) Percentage of pS6 (left) and representative blot of phosphorylation and quantification (right) of the ribosomal protein S6 and ß-actin in control and butyrate macrophages at steady state.(H) Gentamicin protection assay performed on control macrophages, butyrate macrophages, and butyrate macrophages treated for 2 h with the mTOR activator MHY1485 (20 mM).(I) Representative immunoblot of the expression of LC3-II, P62, and ß-actin at steady state or after 2 h infection with Salmonella.(J and K) Protein quantification performed by ImageJ of LC3-II (I) and P62 (J) compared to ß-actin.(L) Degradation of GFP-Salmonella and LC3 induction was assessed by confocal microscopy. Representative images and quantification of GFP fluorescence and LC3 accumulation as outlined in Figure S7C. Data from six independent donors in two independent experiments. Scale bar 5 µm. Each dot represents one donor.Statistical significance was determined using Mann-Whitney U test *p < 0.05, **p < 0.01, and ***p < 0.001.Please also see Figures S2 and S3.

Fig 3: ANDRO treatment induces recovery of general brain metabolism. Treatment with ANDRO stimulated the enzymatic activity of two checkpoint proteins of glycolysis: HK (A) and Pfk (B). Effects of treatments on the levels of AMPK, an important metabolic sensor of glucose metabolism (C). ATP (D) and the ATP/ADP ratio (E) were measured to correlate the previous parameters with the energy status of cells. ANDRO restored the levels of both parameters in J20 mice. Data represent the mean ± SEM of n = 3 (independent experiments), each performed in triplicate. p = 0.05 (95% CI) was considered the threshold for statistical significance, Bonferroni test, p-values are indicated in the “Results” Section.

Fig 4: Schematic model for the action mechanism of lithium. Once internalized, lithium results in the activation of the canonical Wnt signaling pathway, promoting AMPK activation. Both AMPK and the Wnt pathway activation can trigger the upregulation of neuronal Glut3, resulting in glucose metabolism restoration by the stimulation of glucose uptake, increased glycolysis, and elevated ATP levels. Dashed lines indicate the participation of several proteins.

Fig 5: Total, nuclear, and mRNA levels of Nrf2 (A–C), and levels of heme oxygenase-1 (HO-1) (D) in the left ventricles (LVs) of all rat groups. Data were analyzed with one-way ANOVA followed by Tukey’s test. Values are presented as mean ± SD (n = 8/group). Data were considered significantly different at p < 0.5. a: significantly different as compared to control rats; b: significantly different as compared to AKBA-treated rats; c: significantly different as compared to STZ-diabetic rats; d: significantly different as compared to STZ + AKBA-treated rats. CC–compound C (selective AMPK inhibitor).