Fig. 2

Bicyclol did not induce apoptosis or necrosis in HepG2 cells. a The percent of apoptotic and the necrotic cells after 24 h of treatment with different concentrations of bicyclol were measured by flow cytometry. H₂O₂-treated (10 μM) cells were used as positive controls. b Living cell number after co- treatment with bicyclol and z-vad. HepG2 cells were treated with 20 μM z-vad and 500 μM bicyclol at the same time. The cells treated with either 20 μM z-vad or 200 μM bicyclol were used as controls. After a 24 h exposure, the cells were incubated with MTT and the A₅₇₀ was measured. c Flow cytometry analysis of cancer cell apoptosis using the Annexin V-FITC/PI dual-labeling technique. The B2 gate (Annexin V⁺/PI⁺) represents the percentage of necrotic cells, while the B4 gate (Annexin V⁺/PI⁻) represents the percentage of apoptotic cells. Up to 10,000 cells were counted in each sample. d The percent of cells identified by flow cytometry. e The protein level of cleaved caspase-3 treated by bicyclol and Sorafenib