Figure 3.

The apoptotic intrinsic pathway is triggered by 5,6-ECs treatment in HMCLs. (A) JJN3 and U266 cells were seeded in 24-well plates at a density of 2 × 10⁵ cells/well for 24 h and treated with 5,6 α-EC or 5,6 β-EC (20–80 μg/mL) for 24–72 h. Caspase 3/7 enzymatic activity was detected by flow cytometry in both cell lines. The percentage of cells with activated caspase 3/7 was presented in histograms as the mean ± SD (Table S6). The percentages of DiOC₆-(3) negative (with depolarized mitochondria) JJN3 (B) or U266 (C) cells obtained by flow cytometry are showed in histograms as the means ± SD. H₂O₂ (500 µM) was used as a positive control. No statistically significant difference between control and vehicle (EtOH) was noticed. * p < 0.05; ** p < 0.01; *** p < 0.001 with the t-test (Table S7).