Fig. 2: TXNIP induces metabolic reprogramming by inhibiting glycolysis in androgen-dependent PCa cells. | Cell Death & Disease

Fig. 2: TXNIP induces metabolic reprogramming by inhibiting glycolysis in androgen-dependent PCa cells.

From: TXNIP upregulation controls metabolism and cell cycle during androgen deprivation therapy in prostate cancer

Fig. 2

A Representative immunoblotting showing TXNIP overexpression in transfected LNCaP TXNIP cells compared to control LNCaP Empty cells. On the right, representative bright-field micrographs of LNCaP Empty and LNCaP TXNIP cell lines. 20x magnification, scale bar 50 μm. B Proton Efflux Rate (PER) was measured following sequential injection of Rotenone/Animycin A and 2-deoxyglucose (2-DG) as indicated. C Glycolysis-derived Proton Efflux Rate (glycoPER) in basal conditions of LNCaP Empty and LNCaP TXNIP cells and in LNCaP DMSO and LNCaP 20 μM CDX (ADT). D Percentage of protons released due to glycolysis that represent the measured PER of LNCaP Empty, LNCaP TXNIP, LNCaP DMSO and LNCaP 20 μM CDX cells. E GlycoPER from compensatory glycolysis, after cells (LNCaP Empty, LNCaP TXNIP, LNCaP DMSO, and LNCaP 20 μM CDX) are challenged with Rotenone/Antimycin (A). B, C LNCaP TXNIP normalized to LNCaP Empty, and LNCaP CDX normalized to LNCaP DMSO. N = 3 from a representative experiment of three biological replicates. F Extracellular lactate measured in the culture media of both cell lines. B–F measures are normalized against number of cells by DNA-content measurement with Hoescht. N = 3. Data represented as mean ± SEM of a representative experiment from three different experiments. G Histograms of red/green fluorescence determined by JC1 probe measurement to assess mitochondrial membrane potential (MMP). H Ratio red/green was normalized versus Empty group in each of the three different experiments, with N = 3 replicates. Data as mean ± SEM of three different experiments. I Cytoplasmic GLUT1 determination in LNCaP Empty and LNCaP TXNIP. N = 3. J representative images of GLUT1 staining (GLUT1 in green, DAPI in blue). Left and centre panels 63x magnification, scale bar 25 μm. Right panels show in detail the area marked in central panels. White arrows point the main accumulations of GLUT1 in the cells. 150x magnification, scale bar 7.5 μm. For C–I unpaired T-test was performed *p-value < 0.05; **p-value < 0.01; ***p-value < 0.001; ****p-value < 0.0001.

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