Figure 2.

(a) Total apoptosis and necrosis in CRC cells after 4 h of incubation with CuE. Results were expressed as a percentage of the total number of apoptotic cells (early and late apoptosis); (b) the level of intracellular ROS; and (c) the total ROS/Superoxide was detected by flow cytometry.(d) Arrest of cell cycle progression during the G2/M phase in response to CuE treatment. The cell cycle distribution of CRC cell lines was assessed by flow cytometry following staining with PI; (e) results were expressed as a percentage of G2/M; (f) MPM-2 (anti-phospho-Ser/Thr-Pro) expression in untreated and treated cancer cells. MPM-2 is an antibody capable of identifying proteins that are phosphorylated only in mitosis. Cells were dually stained using PI to analyze DNA content, and protein expression was quantified by flow cytometry. As a positive control, separate groups of cells were treated for 24 h with nocodazole (15 μg/ml), an anti-fungal agent known to induce metaphase arrest; (g) cell cycle analysis and the quantification of MPM-2 expression were performed by flow cytometry following treatment with CuE for 24 h. Asterisk (*) in each group of bars indicates that the difference resulting from treatment with CuE 0 μM is statistically significant at P<0.05