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Fig. 4 | Journal of Experimental & Clinical Cancer Research

Fig. 4

From: Ref-1 redox activity alters cancer cell metabolism in pancreatic cancer: exploiting this novel finding as a potential target

Fig. 4

Ref-1 genetic or pharmacological inhibition reduces TCA cycle substrates. Mitochondrial functional assays in Pa03C cells transfected with Scr vs 10 nM siRef-1 (An = 3, *p < 0.05, ##0.0001) and a representative image of the plate. Avg rate of reaction refers to slope of absorbance at 590 vs time. Western blot image of the Pa03C cells after transfection with Ref-1 or Scr siRNA (B). Vinculin is used as the loading control. Average rate of reaction in Pa03C cells treated with Ref-1 redox inhibitor (APX2009, n = 3, *p < 0.05, ##0.0001) or inactive Ref-1 redox inhibitor analog (RN7-58, n = 2) (C) and their representative plate images (D) for 24 h. E Average rate of reaction in CAF02 cells treated with 5 µM APX2009 for 24 h (n = 3). F Fold change of the ratio of NADPH/NADP + in Pa03C cells treated with APX2009 (20 µM) or RN7-58 (20 µM) (n > 2, ##p < 0.0001). G Boxplots show estimated flux of NADP + consuming reactions in one metabolic module using scRNA seq data from Scr vs siRef-1. The two metabolite names on top of each plot are the input and output of each metabolic module. H Measurement of ATP levels by CellTiter-Glo Luminescent Cell Viability Assay in OXPHOS proficient cells (143B WT) or treated with 300 μM phenformin and OXPHOS deficient cells (143B CytB) after treatment with Ref-1 inhibitor APX2009 at the indicated concentrations for 24 h (**p < 0.01, ##0.0001). All data represent Mean ± SE

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