Journal: Journal of Translational Medicine

Article Title: Metabolic reprogramming of poly(morpho)nuclear giant cells determines glioblastoma recovery from doxorubicin-induced stress

doi: 10.1186/s12967-024-05541-9

Figure Lengend Snippet: DOX-induced metabolic reprogramming of T98G cells. ( a , c , g ) Up-regulated β-oxidation ( a ), Krebs cycle proteins ( c ) and mitochondrial respiration proteins ( g ; cf. Fig. for their STRING-generated interactomes) revealed by LC-MS/MS proteomic approach. ( b , f ) Intracellular NADH/NAD ( b ) and ATP/ADP levels in T98G cells ( f ) analysed at the indicated time-points following a pulse DOX treatment. ( d ) The effect of oligomycin A (OliA; 5µM), etomoxir (ETX; 250 µM) and 6-ANA (5µM) on the metabolic activity (left) and PGC formation (right) in T98G populations following a pulse DOX-treatment. ( e ) Quantification of intracellular NADP/NADPH levels in T98G cells at 14th day following a pulse DOX treatment. Statistical significance of the differences was calculated with ANOVA permutation based FDR < 0.05 and post hoc Tukey’s test ( a , c , g ), ANOVA and Tukey’s post hoc test ( d , left) and t-student test ( b ; d ,right, e , f ), * p < 0.05 vs. control. Bars represent SD values. Data representative for >50 cells or 3 independent biological replicates. Note the sensitivity of PGC program to PPP and OXPHOS inhibitors, accompanied by a prominent NADH/ATP and negligible NADPH accumulation in PGCs

Article Snippet: Where indicated, the cells were treated with 3 µM Cis-Platin (Cis-Pt, Sigma; No. P4394; from 10 mM stock in DMSO), 10 µM paclitaxel (PTX; Sigma; No. T7402; from 10 mM stock in DMSO), 250 µM carmustine (Sigma; No. C0400; from 250 mM stock in DMSO), 5 µM 5-fluorouracil (5-FU, Sigma; No. F6627; from 5 mM stock in DMSO), 10–50 µM TWS119 (GSK-3β inhibitor, Sigma; No. SML1271; from 25 mM stocks in DMSO), 5 µM Oligomycin A (OliA, ATP synthase inhibitor, Sigma; No. 7535, from 5 mM stock in DMSO), 250 µM Etomoxir (ETX, β-oxidation inhibitor; Sigma; No. 236020, 2023 from 250 mM stock in DMSO), 5 µM 6-aminonicotinamide (6-ANA, pentose-phosphate pathway inhibitor; from 100 mM stock DMSO), 250–1000 µM sodium ascorbate (Asc, Sigma; No. A4034, from 500 mM stock in PBS), 1–5 mM N-acetyl-L-cysteine (NAC, Sigma; No. A9165; from 5 M stock in PBS), 1–25 mM Taurine (Tau, Sigma; No. T8691; from 5 mM stock in PBS) and 10-1000 µM hydrogen peroxide (H 2 O 2 , POCH; No. 885193111; from freshly prepared 100 mM stock in culture medium).

Techniques: Generated, Liquid Chromatography with Mass Spectroscopy, Activity Assay, Control