Fig 2: Inhibition of FAO decreases ibrutinib-induced cytotoxicity in CLL primary lymphocytes. (A) Survival fraction (relative to non-treated control) after 48 h treatment with ibrutinib alone or in combination with Fatty acids (FA), (n = 10). (B) Survival fraction (relative to non-treated control) after 48 h treatment with etomoxir alone or in combination with ibrutinib, (n = 12). (C) Total glutathione levels after 24 h treatment with etomoxir alone or in combination with ibrutinib (n = 11). Protein expression levels (relative to actin) after 24 h treatment with etomoxir alone or in combination with ibrutinib (D) GS (n = 11), (E) GAC (n = 10), (I) GLUT1 (n=10). (F) NADPH/NADP ratio (n = 8), and (G) NADH/NAD ratio (n = 11) after 24 h treatment with etomoxir alone or in combination with ibrutinib. (H) Relative ROS levels after 48 h treatment with etomoxir alone or in combination with ibrutinib (n = 12), (mean ± SEM). (J) Glucose uptake after 24 h treatment with ibrutinib and etomoxir (n = 8). *p < 0.05, **p < 0.001.

Fig 3: Up-regulation of GLS1 in focal cerebral ischemic brain. (A–C) Representative blot and quantification of GLS1 protein expression levels in ischemic infarction (A), ischemic penumbra (B), and hippocampus (C). (D,E) Focal cerebral ischemic brains and their sham controls were removed after intracranial perfusion and prepared for immunofluorescence staining. Representative pictures of GLS1 and Iba1 immunoreactivities in the hippocampus (D) and cortex (E) of focal cerebral ischemic rat brain sections and control rat brain sections were shown. Images at the lower panels were high-magnification images of the corresponding small box area from the upper panels. Quantification of Iba1+/GLS1+ cells among total cells or Iba1+ cells co-expressing GLS1 immunoreactivities was provided on the right panel. Scale bar: 100 µm. Data were normalized to ß-actin and presented as fold change compared with those in sham rat brain. Error bars denote s.d. from triplicate measurements. *p < 0.05, ***p < 0.001, and ****p < 0.0001 by two-tailed t-test (n = 9).

Fig 4: Simplified scheme of the metabolic pathways analyzed in papillary (PRCC) and clear cell renal cell carcinoma (CCRCC). Red and blue arrows on left and right side of black ones indicate the characteristic rewiring of metabolic pathways in PRCCs and in CCRCCs, respectively. High expressions of certain enzymes suggest balanced nutrient utilization, intensive glutaminolysis (GLS), acetate utilization (ACSS2), fatty acid oxidation (CPT1A), and oxidative phosphorylation (ATPB) were detected in PRCCs (thick red arrows), while high expressions of glycolytic enzymes (GLUT1, HXK2, PFKP) could correlate with higher glycolytic capacity in CCRCCs (thick blue arrows). Created with BioRender.com.

Fig 5: WES analysis of mTOR activity, metabolic enzyme expressions, and in situ mitochondrial stainings of papillary (ACHN) and clear cell (786-O) renal cell carcinoma cell lines. (A) WES analysis of mTOR pathway and metabolic markers in papillary (ACHN), clear cell renal cell carcinoma (786-O), and normal tubular epithelial (HK-2) cell lines. ß-actin was used as loading control. GLS levels were analyzed by a dual-specificity antibody showing both the KGA (60 kDa) and GAC (55 kDa) splice variants. Predicted protein sizes are provided in Table 2. (B) Higher staining intensity with Mitotracker, anti-ATPB, anti-COX IV in ACHN (PRCC) than 786-O (CCRCC) cell lines suggest pronounced mitochondrial OXPHOS function in PRCC cells; however, TOM20 stainings showed no differences (magnification 63×).