Fig. 5
Effect of polyphenols and glycolytic inhibitors on intracellular ROS and GSH levels. a Intracellular accumulation of ROS in HL60 cells, as determined by flow cytometry using H2DCFDA, upon treatment with the indicated concentrations of Quer, Cur, Gen and Lon (μM) or 2-DG (mM), alone and in combination. The results in the bar chart are expressed in relation to untreated (Cont) cells. Cytometry histograms corresponding to untreated cells and cells incubated for 3 h with Quer plus 2-DG are presented as examples. b Intracellular GSH levels in HL60 cells, as determined at 6 h of treatment by monochlorobimane derivatization in luminometric assays. 3-BrP (60 μM) was used as a positive control (see Ref. [44]). The results are expressed in relation to untreated (Cont) cells (approximate GSH content, 8.5 nmol/106 cells). c Frequency of apoptosis inHL60 cells upon incubation for 24 h with 1 mM BSO, alone and in combination with 2-DG or Lon. The combinations Quer plus 2-DG and Quer plus Lon are included as positive controls. Symbols in (a, b) indicate significant differences in relation to Cont, and in (c) between the indicated pairs of values (n.s. non significant). Other conditions, including pre-incubation with Quer in the combined treatments (a, b) and symbols used in statistical analysis, were as in Figs. 1 and 3