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The top 100 differentially expressed proteins, visualized through unsupervised hierarchical clustering (Fig

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1) Oxidative block irreversible, makes NADPH and CO Step 1: G6P + NADP 6-phosphogluconolactone + NADPH via G6PD (rate-limiting) Step 2: lactone + HO 6-phosphogluconate Step 3: 6-PG + NADP ribulose-5-P (Ru5P) + CO + NADPH Net per G6P: 2 NADPH + CO + Ru5P 2) Non-oxidative block reversible sugar shuffling Ru5P ribose-5-P (R5P) or xylulose-5-P (Xu5P) Transketolase (TPP-dependent) moves 2-carbon pieces: Xu5P + R5P sedoheptulose-7-P + GAP

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Collectively, these results show that pharmacological HDM2 inhibition robustly delays the onset of ferroptosis in HT-1080 cells through an on-target mechanism