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Figure 3: Aspirin pretreatment is important for reduction of its toxicity. Cell death in Hek-293 cells, pretreated with 50 μM of aspirin (aspirin pretreated cells) or with vehicle (control cells) for 48 h. In both conditions, toxicity was evaluated in untreated and aspirin treated (5 mM) cells 24 h later. (a) Expression of “trypan blue-not excluding” cells is evaluated as percentage of dead cells in aspirin (5 mM) treated cells compared with control cells. Data were reported as mean ± SD of 3 experiments (NS: not significant). (b) Dead cell profile, representative flow cytometry histograms of Hek-293 cells () stained with 7-AAD, out of three experiments performed. Left and right panels represent control and aspirin pretreated cells, respectively. Percentages of 7-AAD+ cells are indicated in each panel. (c) Ratio of the total numbers of 7-AAD+ cells in 5 mM aspirin treated versus matched untreated cells, both in control and in aspirin pretreated cells. Data were reported as mean ± SD of 3 experiments performed; indicates significant differences in aspirin samples of control versus pretreated cells.