Fig. 4

YGJDSJ increased ROS level in Bel-7402 cells. After 24 h YGJDSJ (100–400 μg/ml) treatment, intracellular ROS production in suspension-cultured Bel-7402 cells was stained with DCFH-DA, observed under fluorescence microscope (× 200) (a), quantitated with a fluorescence microplate reader and expressed as fold of control (b)

Suspension-cultured Bel-7402 cells were pretreated with NAC (50 mmol/L for 2 h) for ROS inhibition, followed by YGJDSJ (100–400 μg/ml) treatment for 24 h, and subjected to caspase-3 (c), caspase-8 (d) and caspase-9 (e) activities and anoikis detection (f). Caspases activities were expressed as fold activation over control. Data shown are representative of three independent experiments. *P < 0.01, versus control group; ^#P < 0.01, versus corresponding dose of YGJDSJ treated NAC (−) group.