Fig. 4.
Ascorbic acid depletes intracellular GSH and increases H2O2 production in MM cell lines.
(A) Cells (4 × 106) were cultured in the absence (■) or the presence of AA (100 μM; ░). GSH levels were determined using the Glutathione Assay kit (Calbiochem) and were normalized to total cellular protein content. Data are presented as mean ± SD of at least 3 independent experiments per cell line. *Means of the AA-treated cells are lower than those of the control cells (P < .001). (B) Cells (2.5 × 105) were cultured for 24 hours in the absence (dashed line) or the presence of AA (100 μM; solid line). Cells were incubated for 30 minutes in 0.5 μM H2DCFDA, washed, and acquired by flow cytometry. Data are representative of at least 3 experiments for each cell line. Percentage cell viability was monitored by PI exclusion and is shown in the inset.