Fig. 5.

Silencing Trx1 or Grx1 affects glycolytic enzymes redox state and activities and glycolytic flux. A) Enzymes of Glycolysis and related pathways undergoing significant (q≤0.1) cysteine redox changes on NOS3 overexpression and upon Trx1 or Grx1 down-regulation have been selected. The precise cysteine residues affected and the changes in the values of their reduced/oxidized cysteine ratios relative to the control (4TO-NOS vs 4TO; siRNA treated vs untreated) are shown. The values for siRNA treated 4TO-NOS cells have been weighted by subtracting the changes observed in siRNA treated 4TO control cells; B) Glycolysis centered metabolic network showing the main glycolytic stream and off-shooting pathways; the final destinations of metabolic fluxes are indicated with thick arrows; the enzymes with redox altered cysteines and their residue position are highlighted. C) Enzymatic activities of triosephosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate kinase, Glycolysis and Pentose Phosphate Pathway fluxes and lactate concentration in siRNA Trx1 and siRNAGrx1 treated 4TO_NOS and 4TO cells are shown relative to untreated 4TO control cells. Between 4 and 9 independent experiments were done for each parameter and a Student t-test was calculated for statistical significance with values p ≤ 0.001, 0.001 >p ≤ 0.01 and 0.01 >p ≤ 0.05 marked with *** , * * and * , respectively.