FIGURE 6.

Baicalein increases OHT‐induced mitochondria‐mediated apoptosis in TAM‐resistant cells. Cells were treated with or without OHT (1 μM) in the presence of baicalein (25 μM) for 48 h. (A) Mitochondrial and intracellular ROS levels were evaluated using MitoSOX and H2DCF‐DA labelling, respectively, with detection via flow cytometry (n = 3). (B) The MMP was measured using TMRE staining and flow cytometry (n = 3). Cells were pretreated with the ROS scavenger NAC (10 mM) for 1 h and then treated with OHT (1 μM) in the presence of baicalein (25 μM) for a further 48 h. (C) Cell apoptosis was detected using annexin V and PI staining coupled with flow cytometry (n = 3). (D) Levels of cytochrome c (Cyt C) in mitochondria or cytoplasm, full‐length caspase‐9 and ‐3 and cleaved caspase‐9 and ‐3 were detected using western blotting. TOM20 and β‐actin were used as the loading control for mitochondrial fractions and cytosolic lysates, respectively. Results are presented as mean ± SD. *p < 0.05, **p < 0.01, control vs. baicalein (Bai); control vs. OHT; OHT vs. OHT plus baicalein; baicalein vs. OHT plus baicalein; baicalein vs. baicalein plus NAC; OHT plus baicalein vs. OHT plus baicalein plus NAC